SMART MODE:

NORMAL GENOMIC

• Ramshaw JA, Shah NK, Brodsky B
• Gly-X-Y tripeptide frequencies in collagen: a context for host-guest triple-helical peptides.
• J Struct Biol. 1998; 122: 86-91
• Display abstract

• The collagen triple-helix consists of a repeating (Gly-X-Y)n sequence. In theory, there are more than 400 possible Gly-X-Y triplets, but analysis of sequences from fibrillar and nonfibrillar collagens shows that only a limited set of triplets are found in significant numbers, and many are never observed. The nonrandom frequency of Gly-X-Y triplets makes it practical to experimentally approach the stability of much of the collagen sequence through the study of a limited set of host-guest peptides. In these peptides, individual Gly-X-Y triplets constitute the guest, while the host consists of Gly-Pro-Hyp tripeptides. A set of host-guest peptides was designed to contain the most common nonpolar and charged triplets found in collagen. All formed stable triple-helices, with their melting temperature depending on the identity of the guest triplet. While including less than 10% of all possible triplets, the data set covers 50-60% of collagen sequences and provides a starting point for establishing a stability scale to predict the relative stability of important collagen regions, such as the matrix metalloproteinase cleavage site or binding sites.

• Sicot FX, Exposito JY, Masselot M, Garrone R, Deutsch J, Gaill F
• Cloning of an annelid fibrillar-collagen gene and phylogenetic analysis of vertebrate and invertebrate collagens.
• Eur J Biochem. 1997; 246: 50-8
• Display abstract

• Arenicola marina possesses cuticular and interstitial collagens, which are mostly synthesised by its epidermis. A cDNA library was constructed from the body wall. This annelid cDNA library was screened with a sea-urchin-collagen cDNA probe, and several overlapping clones were isolated. Nucleotide sequencing of these clones revealed an open reading frame of 2052 nucleotides. The translation product exhibits a triple helical domain of 138 Gly-Xaa-Yaa repeats followed by a 269-residue-long C-terminal non-collagenous domain (C-propeptide). The triple helical domain exhibits an imperfection that has been previously described in a peptide produced by cyanogen bromide digestion (CNBr peptide) of A. marina interstitial collagen. This imperfection occurs at the same place in the interstitial collagen of the vestimentiferan Riftia pachyptila. This identifies the clone as coding for the C-terminal part of a fibrillar collagen chain. It was called FAm1alpha, for fibrillar collagen 1alpha chain of A. marina. The non-collagenous domain possesses a structure similar to carboxy-terminal propeptides of fibrillar pro-alpha chains. Only six conserved cysteine residues are observed in A. marina compared with seven or eight in all other known C-propeptides. This provides information on the importance of disulfide bonds in C-propeptide interactions and in the collagen-assembly process. Phylogenetic studies indicate that the fibrillar collagen 1alpha chain of A. marina is homologous to the R. pachyptila interstitial collagen and that the FAm1alpha gene evolved independently from the other alpha-chain genes. Complementary analyses indicate that the vertebrate fibrillar collagen family is composed of two monophyletic subgroups with a specific position of the collagen type-V chains.

• Yang W, Battineni ML, Brodsky B
• Amino acid sequence environment modulates the disruption by osteogenesis imperfecta glycine substitutions in collagen-like peptides.
• Biochemistry. 1997; 36: 6930-5
• Display abstract

• Ostoegenesis imperfecta (OI) or "brittle bone" disease is associated with mutations in the genes for type I collagen chains and produces variable phenotypes, ranging from lethal cases at birth to mild cases with increased bone fractures. The most common OI mutations are single base substitutions leading to replacement of Gly by another residue, breaking the typical (Gly-X-Y)n repeating sequence pattern of the collagen triple-helix. Triple-helical peptides were designed to focus on residues 892-921 of the alpha1 chain of type I collagen, where two OI Gly-->Ser mutations are found in close proximity, a mild mutation at site 901 and a lethal mutation at site 913. Peptides were designed to include amino acid sequences around these mutation sites, and were synthesized with the normal sequence or with the Gly-->Ser mutated sequence. The peptide including the normal sequence residues 892-909 with four Gly-Pro-Hyp triplets at the C-terminus formed a stable triple-helix, and introduction of a Ser residue for Gly at the 901 mutation site led to a 50% loss of triple-helix content and a decrease in thermal stability, with little effect on folding. A peptide including residues 904-921 again formed a stable triple-helix, but the introduction of the Gly-->Ser substitution at site 913 led to a much greater decrease in thermal stability. These studies demonstrate the impact of local sequences flanking the Gly substitution on structural consequences and support the concept of variability and regional effects along the collagen molecule.

• Kadler K
• Extracellular matrix 1: Fibril-forming collagens.
• Protein Profile. 1995; 2: 491-619

• Tryggvason K
• Molecular properties and diseases of collagens.
• Kidney Int Suppl. 1995; 49: 248-248

• Glumoff V, Makela JK, Vuorio E
• Cloning of cDNA for rat pro alpha 1(III) collagen mRNA. Different expression patterns of type I and type III collagen and fibronectin genes in experimental granulation tissue.
• Biochim Biophys Acta. 1994; 1217: 41-8
• Display abstract

• A cDNA clone for rat pro alpha 1(III) collagen mRNA was isolated from a cDNA library constructed for poly(A)+ RNA from 15-day experimental granulation tissue. Two clones, pRGR1 and pRGR5, were characterized by restriction mapping and sequencing. Comparison with human type III collagen sequences revealed 92% identity at the level of translated amino acids, and 88% identity at nucleotide level in the coding region. In the 3'-untranslated sequence the identity was even higher (90%). The clones were used together with cDNA clones for type I collagen chains, fibronectin and gamma-actin to study the expression of the corresponding mRNAs during the development of experimental sponge-induced granulation tissue in rats. These studies revealed a marked activation of type I and type III collagen genes during the second week of granuloma development followed by a transient reduction in their levels during the third week. The mRNA levels for both collagen types remained relatively unchanged from day 25. The molar ratio of pro alpha 1(III) and pro alpha 1(I) collagen mRNAs was at a maximum on day 6, and then decreased to reach a plateau by the end of the third week. Fibronectin mRNA levels were found to increase slower; the maximum value was reached during the fifth week of granuloma development. The mRNA levels of gamma-actin increased continuously up to the end of the fourth week, thus following the cellular maturation of the tissue.

• Muragaki Y, Abe N, Ninomiya Y, Olsen BR, Ooshima A
• The human alpha 1(XV) collagen chain contains a large amino-terminal non-triple helical domain with a tandem repeat structure and homology to alpha 1(XVIII) collagen.
• J Biol Chem. 1994; 269: 4042-6
• Display abstract

• We have cloned and characterized cDNAs encoding the alpha 1 chain of type XV collagen from a human placenta library. Using primer extension cloning we extended the cDNAs to the 5' end of the mRNA and determined the complete deduced primary structure of the human alpha 1(XV) chain. The polypeptide chain contains nine triple helical domains separated by eight non-triple helical regions and flanked by large amino-terminal (555 amino acid residues) and carboxyl-terminal (256 amino acid residues) non-triple helical domains. Comparison of amino acid sequences of the human alpha 1(XV) chain with those of mouse alpha 1(XVIII) collagen showed remarkable similarity within both amino- and carboxyl-terminal non-triple helical domains. Within the carboxyl third of the amino-terminal domain a tandem repeat structure is found with an about 45-amino acid residue sequence repeated four times. This amino acid sequence has a strikingly high similarity to rat cartilage proteoglycan core protein. Northern blot analysis of human embryonic RNA revealed that alpha 1(XV) mRNA is expressed predominantly in internal organs such as the adrenal gland, kidney, and pancreas.

• Truter S, Andrikopoulos K, Di Liberto M, Womack L, Ramirez F
• Pro-alpha 2(V) collagen gene; pairwise analysis of the amino-propeptide coding domain, and cross-species comparison of the promoter sequence.
• Connect Tissue Res. 1993; 29: 51-9
• Display abstract

• Type V collagen is a minor represented and poorly characterized fibrillar collagen type. Previous cDNA cloning experiments showed that the amino-propeptide of the pro-alpha 2(V) chain shares structural features in common with pro-alpha 1(I), pro alpha 1(II) and pro-alpha 1(III) collagens. In the present paper, this analysis was extended to the gene level. Accordingly the exon/intron arrangement of the amino-propeptide coding domain was compared among pro-alpha 1(I) (COL1A1), pro alpha 1(II) (COL2A1), pro-alpha 1(III) (CO13A1) and pro-alpha 2(V) (COL5A2) collagen genes. This revealed that COL3A1 is the most closely related gene to COL5A2. Based on the assumption that critical regulatory elements might be phylogenetically conserved, we also compared the promoter sequences of the mouse and human COL5A2 genes. This revealed the highest level of sequence homology (97%) in a 52-bp segment which was previously shown to be essential in conferring cell type specificity to the human promoter.

• Walchli C, Koller E, Trueb J, Trueb B
• Structural comparison of the chicken genes for alpha 1(VI) and alpha 2(VI) collagen.
• Eur J Biochem. 1992; 205: 583-9
• Display abstract

• The chicken alpha 1(VI) polypeptide is encoded by a single gene spanning 21 kbp of genomic DNA. This gene is composed of 34 exons and 33 introns. Its structure is closely related to that of the alpha 2(VI) collagen gene, suggesting that the two genes evolved by gene duplication. Both genes contain 19 exons coding for the triple-helical domain. These exons are multiples of 9 bp (27, 36, 45, 54, 63 and 90 bp) and encode an integral number of collagenous Gly-Xaa-Yaa triplets. Since there is no convincing correlation to a building block of 54 bp, it is unlikely that type VI collagen has evolved from a primordial 54-bp module as suggested for all fibrillar collagens.

• Saitta B et al.
• The exon organization of the triple-helical coding regions of the human alpha 1(VI) and alpha 2(VI) collagen genes is highly similar.
• Genomics. 1991; 11: 145-53
• Display abstract

• The alpha 1(VI) and alpha 2(VI) chains, two of the three constituent chains of type VI collagen, are highly similar in size and domain structure. They are encoded by single-copy genes residing in close proximity on human chromosome 21. To study the evolution of the type VI collagen genes, we have isolated and characterized genomic clones coding for the triple-helical domains of the human alpha 1(VI) and alpha 2(VI) chains, which consist of 336 and 335 amino acid residues, respectively. Nucleotide sequencing indicates that, in both genes, the exons are multiples of 9 bp in length (including 27, 36, 45, 54, 63, and 90 bp) except for those encoding for regions with triple-helical interruptions. In addition, the introns are positioned between complete codons. The most predominant exon size is 63 bp, instead of 54 bp as seen in the fibrillar collagen genes. Of particular interest is the finding that the exon structures of the alpha 1(VI) and alpha 2(VI) genes are almost identical. A significant deviation is that a segment of 30 amino acid residues is encoded by two exons of 54 and 36 bp in the alpha 1(VI) gene, but by a single exon of 90 bp in the alpha 2(VI) gene. The exon arrangement therefore provides further evidence that the two genes have evolved from tandem gene duplication. Furthermore, comparison with the previously reported gene structure of the chick alpha 2(VI) chain indicates that the exon structure for the triple-helical domain of the alpha 2(VI) collagen is strictly conserved between human and chicken.

• Hayman AR, Koppel J, Winterhalter KH, Trueb B
• The triple-helical domain of alpha 2(VI) collagen is encoded by 19 short exons that are multiples of 9 base pairs.
• J Biol Chem. 1990; 265: 9864-8
• Display abstract

• We have analyzed the structure of the gene coding for the alpha 2(VI) subunit of chicken type VI collagen. The triple-helical domain of this polypeptide is encoded by 19 short exons distributed over 10 kilobase pairs of genomic DNA. These exons begin with the codon for glycine and end with the codon for the Y amino acid of the collagenous triplet Gly-X-Y. The sizes of the exons are integral multiples of 9 base pairs (bp) (27, 36, 45, 54, 63, and 90 bp), the predominant one being 63 bp. The organization of this type VI collagen gene is therefore quite different from that of the fibrillar collagen genes which have evolved by duplication of a primordial 54-bp unit. It also differs from that of the basement membrane collagen genes whose exon/intron boundaries often split the codons for amino acids.

• Exposito JY, Garrone R
• Characterization of a fibrillar collagen gene in sponges reveals the early evolutionary appearance of two collagen gene families.
• Proc Natl Acad Sci U S A. 1990; 87: 6669-73
• Display abstract

• We have characterized cDNA and genomic clones coding for a sponge collagen. The partial cDNA has an open reading frame encoding 547 amino acid residues. The conceptual translation product contains a probably incomplete triple-helical domain (307 amino acids) with one Gly-Xaa-Yaa-Zaa imperfection in the otherwise perfect Gly-Xaa-Yaa repeats and a carboxyl propeptide (240 amino acids) that includes 7 cysteine residues. Amino acid sequence comparisons indicate that this sponge collagen is homologous to vertebrate and sea urchin fibrillar collagens. Partial characterization of the corresponding gene reveals an intron-exon organization clearly related to the fibrillar collagen gene family. The exons coding for the triple-helical domain are 54 base pairs (bp) or multiples thereof, except for a 57-bp exon containing the Gly-Xaa-Yaa-Zaa coding sequence and for two unusual exons of 126 and 18 bp, respectively. This latter 18-bp exon marks the end of the triple-helical domain, contrary to the other known fibrillar collagen genes that contain exons coding for the junction between the triple-helical domain and the carboxyl propeptide. Compared to other fibrillar collagen genes, the introns are remarkably small. Hybridization to blotted RNAs established that the gene transcript is 4.9 kilobases. Together with previous results that showed the existence of a nonfibrillar collagen in the same species, these data demonstrate that at least two collagen gene families are represented in the most primitive metazoa.

• Soininen R, Huotari M, Ganguly A, Prockop DJ, Tryggvason K
• Structural organization of the gene for the alpha 1 chain of human type IV collagen.
• J Biol Chem. 1989; 264: 13565-71
• Display abstract

• The complete exon size and distribution pattern in the gene for the alpha 1 chain of human type IV collagen was determined. Clones covering 145 kilobases (kb) of genomic DNA including 100 kb of the gene itself as well as 25 kb upstream and 20 kb downstream of the gene sequences, respectively, were isolated from lambda phage and cosmid libraries. The overall gene structure was determined by endonuclease restriction mapping and R-loop analyses and all exon sizes by nucleotide sequencing. The characterized clones contained all the coding sequences except for exon 2 whose sequence was determined after its amplification by the polymerase chain reaction. There were four gaps in the intron sequences; the exact size of the gene is unknown. The entire gene is at least 100 kb in size and contains 52 exons whose size distribution is completely different from that of the genes for fibrillar collagens. In the -Gly-X-Y- coding region there are three exons of 99, 90, and 45 base pairs (bp) each and two exons of 27, 36, 42, 51, 54, 63, and 84 bp each. The rest of the exons have sizes between 71 and 192 bp in the collagenous region. About one-half of the -Gly-X-Y- repeat coding exons start with the second base for the codon of glycine, whereas the other half starts (with two exceptions) with a complete glycine codon. The distribution of split versus unsplit codons is uneven in that the first 19 exons of the gene start with a complete codon. The gene contains repetitive sequences in several regions. A 185-nucleotide segment containing 40 copies of CCT flanked by poly(C) and poly(T) sequences was shown to be located adjacent to an exon. The gene has previously been shown to be located head-to-head to the alpha 2(IV) collagen gene at the distal end of the long arm of chromosome 13, such that the first exons of the two genes are separated by as little as 42 bp (Poschl, E., Pollner, R., and Kuhn, K. (1988) EMBOJ. 7,2687-2695; Soininen, R., Huotari, M., Hostikka, S. L., Prockop, D. J., and Tryggvason, K. (1988) J. Biol. Chem. 263, 17217-17220). The results demonstrate that the human alpha 1(IV) collagen gene has a structure distinctly different from the genes for fibrillar collagens and also that it is considerably larger than any collagen gene characterized to date.

• Su MW, Benson-Chanda V, Vissing H, Ramirez F
• Organization of the exons coding for pro alpha 1(II) collagen N-propeptide confirms a distinct evolutionary history of this domain of the fibrillar collagen genes.
• Genomics. 1989; 4: 438-41
• Display abstract

• The organization of the exons coding for the N-terminal portion of human type II procollagen has been determined. Aside from inferring the previously unknown primary structure of type II N-propeptide, this study has revealed that this coding domain of the gene exhibits an organization uniquely distinct from those of type I and type III collagens. This finding substantiates the notion that the N-propeptide coding domains of the fibrillar collagen genes evolved under less stringent selection than those encoding the C-propeptide and triple helical regions.

• Killen PD, Burbelo P, Sakurai Y, Yamada Y
• Structure of the amino-terminal portion of the murine alpha 1(IV) collagen chain and the corresponding region of the gene.
• J Biol Chem. 1988; 263: 8706-9
• Display abstract

• Collagen IV, the major structural component of basement membranes, is composed of two genetically distinct polypeptide chains, alpha 1(IV) and alpha 2(IV). We have isolated a 522-base-pair (bp) cDNA to the 5' portion of the murine alpha 1(IV) chain mRNA from a library constructed by specific primer extension of poly(A)+ RNA from differentiated F9 cells. This cDNA includes 141 bp of 5' untranslated sequence and encodes a signal peptide plus a portion of the amino-terminal cross-linking (7 S) domain. This cDNA clone was used to obtain the 5' portion of the murine alpha 1(IV) gene from which the nucleotide sequence of exons 1-6 was determined. Exon 1 (234 bp) codes for the 5' untranslated sequence, and the first 28 residues of the protein. The 5' untranslated sequence is highly conserved between the mouse and human species and has the potential to form three mutually exclusive stem-loop structures which may play a role in post-transcriptional regulation. Exons 2-6, which code for the 7 S domain, were found to be 60, 90, 45, and 63 bp in size. The exon structure for the helical portion of the 7 S domain is different from that of the major helical domain, suggesting that they evolved differently.

• Kaytes P, Wood L, Theriault N, Kurkinen M, Vogeli G
• Head-to-head arrangement of murine type IV collagen genes.
• J Biol Chem. 1988; 263: 19274-7
• Display abstract

• The genes for the two dissimilar subunits of type IV collagen are organized in a head-to-head manner with their translation initiation codons within 874 base pairs. Murine genomic clones which contain portions of both genes have been isolated and characterized. These clones contain the first exon of the alpha 1(IV) chain and the first 3 exons for the alpha 2(IV) chain within a 1.7-kilobase HindIII fragment. The intergenic region appears not to resemble previously described bidirectional promoters. The HindIII fragment is present as a single copy in the mouse genome ruling out the presence of one of these gene fragments as a pseudogene. These findings agree with linkage studies of these two genes and differ from the known organization of human fibrillar collagen genes which have been found to be dispersed within the human genome.

• Chu ML et al.
• Amino acid sequence of the triple-helical domain of human collagen type VI.
• J Biol Chem. 1988; 263: 18601-6
• Display abstract

• The complete amino acid sequence of the triple-helical domain of human collagen VI was deduced from sequences of appropriate cDNA clones and confirmed to about 50% by Edman degradation of tryptic peptides. This domain consists of three different peptide segments containing some 335-336 amino acid residues originating from central portions of the alpha 1 (VI), alpha 2(VI), and alpha 3(VI) chains, respectively. Sequence identity in the X/Y positions of the Gly-X-Y repeats is rather low (10-15%) between the chains. Peculiar features of these sequences include 3 cysteine residues about 50 (alpha 3(VI)) and 89 (alpha 1(VI), alpha 2(VI)) residues away from the N-terminus and several Gly-X-Y interruptions clustered in the C-terminal two-thirds of the triple helix. These structures are presumably required for cross-linking collagen VI oligomers and for super-coiling of triple helices in the dimers. Other features include 11 Arg-Gly-Asp sequences, some of which are likely to be used as cell-binding sites, and four Asn-X-Thr sequences, allowing N-linked glycosylation along the triple helix. Junctional areas close to the helix contain short, cysteine-rich segments which may seal the triple-helical domain through disulfide bond formation, endowing it with high stability. These features, together with a low sequence homology to fiber-forming and basement-membrane collagens, document the unique character of collagen VI, whose triple helix is specifically adjusted for forming microfibrils in tissues.

• Bernard M et al.
• Cloning and sequencing of pro-alpha 1 (XI) collagen cDNA demonstrates that type XI belongs to the fibrillar class of collagens and reveals that the expression of the gene is not restricted to cartilagenous tissue.
• J Biol Chem. 1988; 263: 17159-66
• Display abstract

• We have isolated several overlapping cDNA clones encoding alpha 1(XI) collagen chains from human and rat cDNA libraries. Together the human cDNAs code for 335 uninterrupted Gly-X-Y triplets, and a 264-amino acid C-propeptide, while the rat cDNAs cover the entire C-propeptide and about a third of the triple-helical domain. Comparison of the human and rodent nucleotide sequences showed a 95% sequence similarity. The identification of the clones as alpha 1(XI) cDNAs was based on the complete identity between the amino acid sequences of three human alpha 1(XI) cyanogen bromide peptides and the cDNA-derived sequence. Examination of and the cDNA-derived amino acid sequence showed a variety of structural features characteristic of fibrillar-forming collagens. In addition, nucleotide sequence analysis of a selected portion of the corresponding human gene revealed the characteristic 54-base pair exon motif. We conclude therefore that pro-alpha 1 (XI) collagen belongs to the group of fibrillar collagen genes. We also suggest that the expression of this gene is not restricted to cartilage, as previously thought, since the cDNA libraries from which the clones were isolated, originated from both cartilagenous and noncartilaginous tissues.

• McCormick D, van der Rest M, Goodship J, Lozano G, Ninomiya Y, Olsen BR
• Structure of the glycosaminoglycan domain in the type IX collagen-proteoglycan.
• Proc Natl Acad Sci U S A. 1987; 84: 4044-8
• Display abstract

• Type IX collagen represents 5-20% of the total collagen in hyaline cartilage. The molecules of this collagen are composed of three genetically distinct polypeptide subunits. One of these subunits, alpha 2(IX), contains covalently bound glycosaminoglycan (chondroitin sulfate or dermatan sulfate). We report here on the structure of the glycosaminoglycan attachment site of type IX collagen-proteoglycan. We show, by a combination of cDNA and peptide sequencing, that the attachment region contains the sequence Gly-Ser-Ala-Asp, located within the noncollagenous domain NC3 of the alpha 2(IX) chain. By comparing the exons encoding the NC3 domain in the alpha 2(IX) and alpha 1(IX) genes, we find that the exon coding for the glycosaminoglycan attachment site in the alpha 2(IX) gene is 48 base pairs long, whereas the homologous alpha 1(IX) exon is 33 base pairs. The NC3 domain is, therefore, five amino acid residues longer in alpha 2(IX) than in alpha 1(IX). The extra sequence in alpha 2(IX), Val-Glu-Gly-Ser-Ala, provides a simple explanation for the kink observed at the NC3 domain of type IX molecules when examined by electron microscopy. The inserted block of amino acid residues also provides the NC3 domain of alpha 2(IX) chains with a serine residue, not present in alpha 1(IX) that serves as attachment site for a glycosaminoglycan side chain. Our data show that the amino acid sequence that surrounds the glycosylated serine residue in type IX collagen-proteoglycan differs from glycosylated sequences in noncollagenous core proteins. The data also provide strong evidence that glycosylation of type IX collagen is not a chance glycosylation of a serine residue in a noncollagenous domain, but is a specific post-translational modification of this unusual collagen molecule.

• Kurkinen M et al.
• Extensive homology between the carboxyl-terminal peptides of mouse alpha 1(IV) and alpha 2(IV) collagen.
• J Biol Chem. 1987; 262: 8496-9
• Display abstract

• We have determined the complete primary structure for the carboxyl-terminal peptides of mouse alpha 1(IV) and alpha 2(IV) collagen; which have 229 and 227 amino acids, respectively. The amino acid sequences are 63% identical and conservatively substituted in 28 positions. A striking feature of these peptides is that the first half of each sequence is homologous with the second half, 37% in alpha 1(IV) and 36% in alpha 2(IV). These results suggest that the carboxyl-terminal peptides of type IV collagen are closely related in their structure and evolution. Presumably, they were first derived by internal duplication of a common ancestral DNA sequence which later, by gene duplication, gave rise to the two different but homologous carboxyl-terminal peptides of type IV collagen.

• Hostikka SL, Tryggvason K
• Extensive structural differences between genes for the alpha 1 and alpha 2 chains of type IV collagen despite conservation of coding sequences.
• FEBS Lett. 1987; 224: 297-305
• Display abstract

• Analysis of the structure of the 3'-end of the human alpha 2(IV) gene demonstrated that the alpha 1(IV) and alpha 2(IV) genes have diverged extensively in spite of the apparent homology of the respective gene products. The NC-1 domain and the 3'-untranslated region are encoded by three exons in the alpha 2(IV) gene but five exons in the alpha 1(IV) gene. The two introns present in the NC-1 domain coding part of the alpha 2(IV) gene had the same location as two of the introns of the alpha 1(IV) gene. The junction exon in the alpha 2(IV) gene contains 53 bp coding for Gly-X-Y sequences whereas there are 71 bp in the alpha 1(IV) gene. Three other Gly-X-Y coding exons studied from the human alpha 2(IV) gene have sizes that differ from corresponding exons in the alpha 1(IV) gene and only one intron location matches here between the two genes. None of the exons studied has 54 bp or multiples thereof.

• Sakurai Y, Sullivan M, Yamada Y
• Alpha 1 type IV collagen gene evolved differently from fibrillar collagen genes.
• J Biol Chem. 1986; 261: 6654-7
• Display abstract

• Type IV collagen is a major structural component in basement membranes. It is considerably different from the fibrillar collagens, types I-III. For example, unlike fibrillar collagens, the triple helical domain of type IV collagen is frequently interrupted by nonhelical regions. In this report, we demonstrate several overlapping genomic clones which cover most of the mouse alpha 1(IV) chain. Electron microscopic analysis of R-loops revealed that there were at least 28 exons within 35 kilobases of the gene segment. The sizes of six exons were determined by DNA sequence analysis to be 81, 178, 134, 73, 129, and 213 base pairs. These sizes do not appear to be related to the 54-base pair coding unit which is characteristic of fibrillar collagen exons, suggesting that the alpha 1 type IV collagen gene evolved differently from the fibrillar collagen genes.

• Vogeli G, Horn E, Carter J, Kaytes PS
• Proposed alignment of helical interruptions in the two subunits of the basement membrane (type IV) collagen.
• FEBS Lett. 1986; 206: 29-32
• Display abstract

• We have isolated a cDNA clone for part of the alpha 2 type IV collagen (pCIV-2-176). Deoxynucleotide sequence analysis shows that this clone codes for 439 amino acids from the helical domain adjacent to the C-terminal globular domain of the alpha 2 (IV) chain. By aligning the deduced amino acid sequence of the alpha 2 (IV) chain with the published sequence for the alpha 1 (IV) chain, we find that all interruptions in the alpha 1 (IV) chain coincide with an interruption in the alpha 2 (IV) chain. Additional interruptions in the alpha 2 (IV) chain exist, however, three out of the four analysed only slightly disturb the collagen triple helix.

• Miskulin M et al.
• Human type III collagen gene expression is coordinately modulated with the type I collagen genes during fibroblast growth.
• Biochemistry. 1986; 25: 1408-13
• Display abstract

• Type III collagen is one of the major interstitial collagens and, as such, plays an important role in modulating the structure and function of most tissues. To compare the expression of the type III collagen gene to that of the type I collagen alpha 1(I) and alpha 2(I) genes, cDNAs encoding the 3' one-third of the human alpha 1(III) collagen mRNA were obtained by screening a human fetal lung fibroblast cDNA library with a cloned segment of the chicken alpha 1(III) gene. Northern blot analysis of human fetal lung fibroblast RNA demonstrated two alpha 1(III)-specific mRNAs of sizes 6.6 and 5.8 kilobases, sizes clearly different from those of the type I collagen mRNAs. Analyses of populations of dividing and nondividing human lung fibroblasts revealed that, on a per cell basis, the nondividing population contained twice as much alpha 1(III) mRNA than did the dividing population. The same was true for the type I collagen alpha 1(I) and alpha 2(I) mRNA transcripts. Similar results were obtained when alpha 1(III), alpha 1(I), and alpha 2(I) mRNA transcripts were quantified by using dot blot evaluation of total RNA, Northern analysis of total RNA, and dot blot evaluation of cytoplasmic RNA. Thus, despite the fact that the alpha 1(III) collagen gene is located on a chromosome different from the alpha 1(I) and alpha 2(I) genes, the expression of these three collagen chains appears to be coordinately controlled during periods of rapid and slow fibroblast growth.

• Benveniste-Schrode K, Doering JL, Hauck WW, Schrode J, Kendra KL, Drexler BK
• Evolution of chick type I procollagen genes.
• J Mol Evol. 1985; 22: 209-19
• Display abstract

• Although the major types of vertebrate collagen have a number of structural properties in common, significant DNA sequence homologies have not been detected between different portions of the helical coding domains within the same gene or between different genes. However, under non-stringen hybridization conditions we found considerable cross-homology within and between alpha 1(I) and alpha 2(I) chick cDNAs in the coding regions for helical sequences. Detailed analyses at the DNA sequence level have led us to propose that the gene for chick pro alpha 2(I) collagen arose from a 9-bp primordial sequence. A consensus sequence for the 9-bp repeat was derived: GGTCCTCCT, which codes for a Gly-Pro-Pro triplet. The primordial ancestor of this 9-bp unit, GGTCCTXCT, apparently underwent duplication and divergence. Each resulting 9-bp sequence was triplicated to form a 27-bp domain, and a condensation event produced a 54-bp domain. This genetic unit then underwent multiple rounds of amplification to form the ancestral gene for the full-length helical section of alpha 2(I). A different 9-bp consensus sequence (GGTCCCCCC) seems to have been the basis of the chick pro alpha 1(I) gene.

• Lozano G, Ninomiya Y, Thompson H, Olsen BR
• A distinct class of vertebrate collagen genes encodes chicken type IX collagen polypeptides.
• Proc Natl Acad Sci U S A. 1985; 82: 4050-4
• Display abstract

• Type IX collagen is a disulfide-bonded protein first isolated from hyaline cartilage. The structure of this collagen is unusual in that the molecules contain three triple-helical domains interspersed with noncollagenous regions. The molecules are heterotrimers composed of three genetically distinct polypeptide chains. In our laboratory, cDNAs specific for two of these polypeptide chains have recently been isolated. Here we report on the isolation of genomic clones by use of these cDNAs as probes for screening a chicken genomic library. Nucleotide sequence analysis of these clones shows that the exon structure of type IX collagen genes is fundamentally different from the exon structure of the genes for the fibrillar collagen types I-III. Whereas the sizes of exons in fibrillar collagen genes are related to a basic 54-base-pair coding unit, the exons of type IX collagen genes show a large variation in size and do not appear to be related to a 54-base-pair unit. We propose, therefore, that type IX collagen genes belong to a class of vertebrate collagen genes distinct from that of fibrillar collagens.

• van der Rest M, Mayne R, Ninomiya Y, Seidah NG, Chretien M, Olsen BR
• The structure of type IX collagen.
• J Biol Chem. 1985; 260: 220-5
• Display abstract

• We present a detailed analysis both of tryptic peptides and amino-terminal sequences of the subunits of two collagenous fragments (HMW and LMW) previously isolated from pepsin extracts of chicken cartilage (Reese, C.A., and Mayne, R. (1981) Biochemistry 20, 5443-5448). This analysis and a comparison with the nucleotide sequence of the cDNApYN1738 (Ninomiya, Y., and Olsen, B.R. (1984) Proc. Natl. Acad. Sci. U. S. A. 81, 3014-3018) shows that HMW and LMW are pepsin-resistant fragments of a unique collagen composed of molecules with three different polypeptide chains (alpha-chains). This collagen has been assigned the type number IX, and the alpha-chain encoded by pYN1738 has been given the designation alpha 1 (IX). Type IX collagen contains three triple-helical domains and at least two sets of interchain disulfide bridges. At the amino and carboxyl ends are noncollagenous domains which do not appear to be homologous to amino and carboxyl propeptides of interstitial collagens.

• Myers JC, Loidl HR, Stolle CA, Seyer JM
• Partial covalent structure of the human alpha 2 type V collagen chain.
• J Biol Chem. 1985; 260: 5533-41
• Display abstract

• Human cDNA libraries were screened with a cDNA fragment presumably encoding the 3' terminus of a procollagen carboxyl propeptide not identifiable as types I, II, III, or IV by protein sequence or Northern blot hybridization. One clone contained a 1350-base pair insert coding in part for 55 uninterrupted Gly-X-Y triplets. Comparison with the amino acid composition of the COOH-terminal cyanogen bromide (CB) peptides of the alpha 1 and alpha 2 type V collagen chains showed similarity only to the alpha 2(V)CB fragment. To identify the NH2 terminus of the peptide designated by methionine, an additional isolate was sequenced and found to contain a Gly-Met-Pro triplet. Thirty-one amino acids from the NH2 terminus of the alpha 2(V)CB9 fragment were then determined by Edman degradation and found to be identical to those derived from the cDNA clone. The DNA sequence encoding part of the triple helical region establishes for the first time the partial structure of a type V collagen chain. Although comparison of residues 796-1020 of the alpha 2(V) collagenous region with alpha 1 (III), alpha 1(I), and alpha 2(I) shows strong conservation of charged positions, the latter three chains appear considerably more similar to each other than to alpha 2(V). A striking feature of the alpha 2(V) sequence between 918-944 is the absence of proline residues. In the analogous region of alpha 1(I) where this amino acid is also lacking, a flexible site in the rigid triple helical structure of type I collagen has been observed (Hofmann, H., Voss, T., Kuhn, K. and Engel, J. (1984) J. Mol. Biol. 172, 325-343).

• Upholt WB, Strom CM, Sandell LJ
• Structure of the type II collagen gene.
• Ann N Y Acad Sci. 1985; 460: 130-40
• Display abstract

• In summary, the exon/intron structure of the chicken type II collagen gene is identical with that of the chicken alpha 2(I) collagen gene and differs at only one known position from the human and mouse alpha 1(I) genes. However, the chicken type II gene is different from the chicken alpha 2(I) gene in that it is considerably shorter because of a much smaller average intron size and in that the G+C composition of the introns is much higher. The codon usage of the type II genes also shows characteristic differences. There is a single copy of the chick type II gene per haploid genome.

• Kurkinen M, Bernard MP, Barlow DP, Chow LT
• Characterization of 64-, 123- and 182-base-pair exons in the mouse alpha 2(IV) collagen gene.
• Nature. 1985; 317: 177-9
• Display abstract

• Genes encoding types I, II and III collagens (fibrillar collagens) contain many discrete-size exons, most of them 54 base pairs (bp) long, in addition to the 45-, 99-, 108- and 162-bp exons. It has been suggested that these collagen genes evolved from an ancestral coding unit of 54 bp. Type IV collagen is a specific component of basement membranes and contains two genetically distinct polypeptides, the alpha 1(IV) and alpha 2(IV) chains. It differs from the types I-III collagens in that it contains interruptions in the Gly-X-Y repeat sequence and does not form ordered fibrillar structures. We have isolated complementary DNA and genomic clones for the mouse alpha 2(IV) collagen chain and here characterize 64-, 123- and 182-bp exons in the Gly-X-Y coding domain of the gene. The data suggest that the alpha 2(IV) collagen gene may have evolved differently from those encoding the fibrillar collagens.

• French BT, Lee WH, Maul GG
• Nucleotide sequence of a cDNA clone for mouse pro alpha 1(I) collagen protein.
• Gene. 1985; 39: 311-2
• Display abstract

• A cDNA clone for mouse pro alpha 1(I) collagen has been isolated and sequenced. A 1.8-kb PstI fragment spans nucleotides -1991 to -159 of the alpha-domain of mouse pro alpha 1(I) collagen.

• Liau G, Mudryj M, de Crombrugghe B
• Identification of the promoter and first exon of the mouse alpha 1 (III) collagen gene.
• J Biol Chem. 1985; 260: 3773-7
• Display abstract

• We have identified the promoter-proximal exon of the mouse alpha 1 (III) collagen gene using a synthetic oligonucleotide as a hybridization probe and have determined the DNA sequence of this exon and 380 base pairs 5' to it. The exact start site of transcription was localized with a primer extension experiment. The region upstream of the start of transcription shows only scattered homologies with the analogous sequences in the alpha 1(I) and alpha 2(I) mouse collagen genes although these genes are often co-expressed and co-regulated. The most striking homology with the type I gene is seen around the start of translation. This region contains an inverted repeat which could form a stem-loop structure with a calculated delta G of -30 kcal in the type III collagen mRNA. When compared to the alpha 1(I) and alpha 2(I) signal peptides, the signal peptide of mouse alpha 1(III) collagen presents less homology than when these segments are compared to each other.

• Boedtker H, Finer M, Aho S
• The structure of the chicken alpha 2 collagen gene.
• Ann N Y Acad Sci. 1985; 460: 85-116

• Sandell LJ, Prentice HL, Kravis D, Upholt WB
• Structure and sequence of the chicken type II procollagen gene. Characterization of the region encoding the carboxyl-terminal telopeptide and propeptide.
• J Biol Chem. 1984; 259: 7826-34
• Display abstract

• The DNA sequence of two overlapping cDNA clones and a genomic lambda clone covering the region coding for 288 amino acids at the COOH terminus of the chicken type II procollagen gene is reported. This region consists of 4 exons coding for the last 15 amino acids of the triple helical domain and 273 amino acids which correspond to the COOH-terminal telopeptide and COOH-terminal propeptide. The sequence, base composition, and codon usage of this region of the type II procollagen gene show particularly high similarity to those of the chicken alpha 1(I) procollagen gene and differ from those of the alpha 2(I) and alpha 1(III) gene sequences. Two DNA tracts of low sequence similarity were observed. One of these regions spans the telopeptide and COOH-terminal propeptidase cleavage site, although 4-5 amino acids at the actual cleavage site are conserved compared with the alpha 1(I) and alpha 2(I) genes. A region of unusually high nucleotide sequence conservation is present in exon 2 (amino acids 171c - 186c ) consisting of approximately 45 nucleotides with only one or two base substitutions compared with the other procollagen genes. Within this conserved sequence is a site for carbohydrate attachment. The 3' nontranslated sequence of the type II procollagen mRNA is longer than that of either the alpha 1(I) or alpha 2(I) mRNA and contains several unusual long tracts consisting primarily of one or two bases. Although the canonical site for polyadenylation is not present, two related sequences, AACAAA and ATATAAA , are present 32 and 41 bases preceding the end of the major RNA species. The exon/intron structure of the type II procollagen gene is similar to that of other collagen genes which have been described. This DNA sequence provides the first extensive report of the amino acid sequence of chicken type II procollagen.

• Aho S, Tate V, Boedtker H
• Location of the 11 bp exon in the chicken pro alpha 2(I) collagen gene.
• Nucleic Acids Res. 1984; 12: 6117-25
• Display abstract

• During the fine structural analysis of the 5' end of the 38 kb chicken pro alpha 2(I) collagen gene, we failed to locate an exon, only 11 bp in size, which had been predicted from the DNA sequence analysis of a cDNA clone complementary to the 5' end of the pro alpha 2(I) collagen mRNA (1). We know report the location of this 11 bp exon, exon 2, at the 5' end of a 180 bp Pst I fragment, 1900 bp 3' to exon 1 and 600 bp 5' to exon 3. Its sequence, ATGTGAGTGAG, is highly unusual in that it contains two overlapping consensus donor splice sequences. Moreover, it is flanked by two overlapping donor splice sequences but only one of the four splice sequences is actually spliced (1). The first half of intron 1 also has an unusual sequence: it is 68% GC, contains 88 CpG dinucleotides and 11 Hpa II sites. The second half is more like other intron sequences in the collagen gene with a GC content of 41%, 19 CpG, and no Hpa II sites. However it contains two sequences with 7 and 9 bp homology to the 14 bp SV40 enhancer core sequence. It is suggested that some part of intron 1 may be involved in regulation.

• Schmidt A, Yamada Y, de Crombrugghe B
• DNA sequence comparison of the regulatory signals at the 5' end of the mouse and chick alpha 2 type I collagen genes.
• J Biol Chem. 1984; 259: 7411-5
• Display abstract

• We have isolated a DNA segment that contains the presumptive transcriptional and translational control signals located at the 5' end of the mouse alpha 2(I) collagen gene. The DNA sequence of the first exon of this gene and of 400 base pairs preceding the start of transcription was compared with an analogous sequence in the chick alpha 2(I) collagen gene. The most striking similarity is a block of 68 nucleotides surrounding the translation initiation site which is perfectly conserved except for four contiguous nucleotides in the middle of the conserved sequence. These four nonconserved bases lie between the arms of a perfect inverted repeat with nine nucleotides in each arm. The very high degree of sequence conservation of this segment suggests that both the inverted repeat and the rest of the perfectly conserved sequence have a regulatory role, possibly in translation. The segment preceding the transcription start shows blocks of similarities interspersed with dissimilar sequences. The conserved sequences include the TATA box, CAT box, a G/C rich sequence around -110, a segment that contains almost exclusively pyrimidines on one strand and is sensitive to endonuclease S1 in both promoters, and several other segments that extend to 350 base pairs preceding the start of transcription. Although the distances between these regions of similarity and the TATA box are not the same in the two promoters, we believe the conserved sequences play a role in the transcriptional activation of this gene.

• Chu ML et al.
• Human pro alpha 1(I) collagen gene structure reveals evolutionary conservation of a pattern of introns and exons.
• Nature. 1984; 310: 337-40
• Display abstract

• The collagens represent an interesting example of a structurally related but genetically distinct family of proteins. Type I, the most abundant of the vertebrate collagens, comprises two pro alpha 1(I) chains and one pro alpha 2(I) chain, each containing terminal propeptides and a central domain of 338 (Gly, X, Y) repeats. The structure of the chicken pro alpha 2(I) gene shows an intriguing relationship between exon organization and the arrangement of (Gly, X, Y) repeats (see ref. 2 for review). This has led to the suggestion that the collagens evolved from a common ancestral unit of 54 base pairs (bp). Here we present the structure of the entire human pro alpha 1(I) gene and compare this with the chicken pro alpha 2(I). The exon arrangement of the two genes is remarkably similar, although the human pro alpha 1(I) is more compact because of the shorter length of its introns. The data strongly support the notion that the type I genes have evolved from an ancestral multi-exon unit, and that once the gene was translated, a strong evolutionary pressure caused it to maintain this elaborate structure.

• Yamada Y, Mudryj M, Sullivan M, de Crombrugghe B
• Isolation and characterization of a genomic clone encoding chick alpha 1 type III collagen.
• J Biol Chem. 1983; 258: 2758-61
• Display abstract

• We have isolated a cloned segment of the chick alpha 1 type III collagen gene from a genomic DNA library. This DNA clone was obtained by cross-hybridization with a cDNA clone containing sequences coding for a segment of the carboxy propeptide of chick alpha 1 type I collagen. This genomic clone was identified as coding for alpha 1 type III collagen by DNA sequence analysis of two segments of the gene and by its ability to hybridize to crop mRNA but not to calvaria mRNA in agreement with the presence of alpha 1 type III collagen in crop but not in calvaria. The deduced amino acid sequences reveal characteristic features for alpha 1 type III collagen and are in good agreement with the known protein sequences of calf and human alpha 1 type III collagen. The size of alpha 1 type III mRNA is about 6 kilobases, slightly larger than the mRNA for alpha 1 type I. R-loop electron microscopic analysis indicates that at least 16 exons are interspersed in a 9-kilobase DNA segment that contains the third distal part of the alpha 1 type III collagen gene.

• Kimura S
• Vertebrate skin type I collagen: comparison of bony fishes with lamprey and calf.
• Comp Biochem Physiol B. 1983; 74: 525-8
• Display abstract

• 1. Characterization of Type I collagen alpha chains, alpha 1 and alpha 2, in the skin tissues of carp and common mackerel revealed a marked interspecies difference in CNBr-peptide maps of the alpha 2 chains, suggesting the hypervariability during evolution of the alpha 2 chains relative to the alpha 1 chains. 2. When compared with calf Type I collagen, lower vertebrate Type I collagens derived from these bony fishes as well as from lamprey were found to exhibit a higher degree of structural similarity between their alpha 1 and alpha 2 chains.

• Henkel W, Glanville RW
• Covalent crosslinking between molecules of type I and type III collagen. The involvement of the N-terminal, nonhelical regions of the alpha 1 (I) and alpha 1 (III) chains in the formation of intermolecular crosslinks.
• Eur J Biochem. 1982; 122: 205-13

• Scott PG, Pearson H
• Cathepsin D: specificity of peptide-bond cleavage in type-I collagen and effects on type-III collagen and procollagen.
• Eur J Biochem. 1981; 114: 59-62
• Display abstract

• 1. Cathepsin D, purified from bovine thymus, has a limited proteolytic effect on types I and III bovine collagens. The alpha 1 (I) chain was cleaved in native or denatured collagen only within the carboxy-terminal extra-helical sequence, the major site being between resides C6 (Leu) and C7 (Ser). The alpha 2 chain was unaffected in native collagen but was slowly cleaved between residues 782 (Phe) and 783 (Leu) in the denatured form. Cleavages, at 45 degrees C, in type III collagen occur within the extra-helical amino-terminal sequence, on the carboxy-terminal side of the lysine residue involved in intermolecular cross-linking. All three sites of action are within sequences of general hydrophobic character. 2. The very restricted cleavage of peptide bonds in denatured collagens can be ascribed to the infrequent occurrence of groupings of more than two hydrophobic residues and to the high content of the conformation-limiting residues proline and hydroxyproline. 3. The previously demonstrated failure of cathepsin D to solubilize a representative proportion of type III collagen from the fibres of bovine skin collagen [P.G. Scott and C.H. Pearson (1978) Biochem, Soc, Trans. 6, 1197-1199] may be explained by lack of ability of the enzyme to act on this collagen at 25 degrees C, in such a manner as to separate molecules joined by intermolecular cross-links involving the amino-terminal extrahelical region of the molecule.

• Fuller F, Boedtker H
• Sequence determination and analysis of the 3' region of chicken pro-alpha 1(I) and pro-alpha 2(I) collagen messenger ribonucleic acids including the carboxy-terminal propeptide sequences.
• Biochemistry. 1981; 20: 996-1006
• Display abstract

• Three pro-alpha 1 collagen cDNA clones, pCg1, pCg26, and pCg54, and two pro-alpha 2 collagen cDNA clones, pCg 13 and pCg45, were subjected to extensive DNA sequence determination. The combined sequences specified the amino acid sequences for chicken pro-alpha 1 and pro-alpha 2 type I collagens starting at residue 814 in the collagen triple-helical region and continuing to the procollagen C-termini as determined by the first in-phase termination codon. Thus, the sequences of 272 pro-alpha 1 C-terminal, 260 pro-alpha 2 C-terminal, 201 pro-alpha 1 helical, and 201 pro-alpha 2 helical amino acids were established. In addition, the sequences of several hundred nucleotides corresponding to noncoding regions of both procollagen mRNAs were determined. In total, 1589 pro-alpha 1 base pairs and 1691 pro-alpha 2 base pairs were sequenced, corresponding to approximately one-third of the total length of each mRNA. Both procollagen mRNA sequences have a high G+C content. The pro-alpha 1 mRNA is 75% G+C in the helical coding region sequenced and 61% G&C in the C-terminal coding region while the pro-alpha 2 mRNA is 60% and 48% G+C, respectively, in these regions. The dinucleotide sequence pCG occurs at a higher frequence in both sequences than is normally found in vertebrate DNAs and is approximately 5 times more frequent in the pro-alpha 1 sequence than in the pro-alpha 2 sequence. Nucleotide homology in the helical coding regions is very limited given that these sequences code for the repeating Gly-X-Y tripeptide in a region where X and Y residues are 50% conserved. These differences are clearly reflected in the preferred codon usages of the two mRNAs.

• Hofmann H, Fietzek PP, Kuhn K
• Comparative analysis of the sequences of the three collagen chains alpha 1(I), alpha 2 and alpha 1(III) Functional and genetic aspects.
• J Mol Biol. 1980; 141: 293-314

• Seyer JM, Mainardi C, Kang AH
• Covalent structure of collagen: amino acid sequence of alpha 1 (III)-CB5 from type III collagen of human liver.
• Biochemistry. 1980; 19: 1583-9
• Display abstract

• Type III collagen was prepared from human liver by limited pepsin digestion, differential salt precipitation, and carboxymethylcellulose chromatography. Ten distinct peptides were obtained by cyanogen bromide digestion. The peptide alpha 1 (III)-CB5 was further purified by carboxymethylcellulose chromatography, and its amino acid sequence was determined. Automatic Edman degradation of intact alpha 1 (III)-CB5, tryptic and thermolytic peptides, and hydroxylamine-derived fragments was used to establish the total sequence. The mammalian collagenase site contained in the alpha 1 (III)-CB5 sequence was ascertained by digestion of native type III collagen with purified rheumatoid synovial collagenase. Collagenase cleavage occurred at a single Gly--Ile bond, one triplet before the corresponding specific cleavage site of type I collagen. The present work brings the known sequence of human liver type III collagen to include alpha 1 (III)-CB3-7-6-1-8-10-2-4-5. These correspond to the homologous region of alpha 1 (I)-CB0-1-2-4-5-8-3-7 residues 11--804.

• Lindy S, Turto H
• [Degradation of collagen]
• Duodecim. 1980; 96: 1473-9

• Showalter AM et al.
• Nucleotide sequence of a collagen cDNA-fragment coding for the carboxyl end of pro alpha 1(I)-chains.
• FEBS Lett. 1980; 111: 61-5

• Schafer MP, Boyd CD, Tolstoshev P, Crystal RG
• Structural organization of a 17 KB segment of the alpha 2 collagen gene: evaluation by R loop mapping.
• Nucleic Acids Res. 1980; 8: 2241-53
• Display abstract

• A recombinant phage, SpC3, containing a 17 kb genomic DNA insert representing approximately 60% of the 3' portion of the sheep collagen alpha 2 gene, was evaluated by electron microscopic R loop analysis. A minimum of 17 intervening sequences (introns) and 18 alpha 2 coding sequences (exons) were mapped. With the exception of the 850 base pair exon located at the extreme 3' end of the insert, all exons contained 250 base pairs or less. The total length of all the exons in SpC3 was 3,014 base pairs. The length distribution of the 17 introns ranged from 300 to 1600 base pairs; together, all of the introns comprised 14,070 base pairs of SpC3 DNA. Thus, the DNA region required for coding the interspersed 3 kb of alpha 2 collagen genetic information was 5.6 fold longer than the corresponding alpha 2 mRNA coding sequences.

• Vogeli G et al.
• Isolation and characterization of genomic DNA coding for alpha 2 type I collagen.
• Nucleic Acids Res. 1980; 8: 1823-37
• Display abstract

• We have isolated and characterized a segment of the chick alpha 2 collagen gene by screening a library of chick genomic fragments using as hybridization probe an alpha 2 collagen cDNA clone. Several clones were isolated and one of them, lambda gCOL 204, was used for further studies. The DNA of lambda gCOL 204 hybridizes to a unique species of mRNA the size of alpha 2 collagen mRNA. This mRNA can be translated into a unique polypeptide which comigrates in SDS-gel electrophoresis with pro-alpha 2 collagen. Electron microscopic analysis by R-loop technique indicates that lambda gCOL 204 contains 7Kb of the alpha 2 collagen gene. This 7 Kb piece constitutes the 3' end of the gene. The same clone also contains 9 Kb of DNA that is immediately adjacent to the 3' end of the alpha 2 collagen gene. The cloned segment of the alpha 2 collagen gene is interrupted by 8 intervening sequences of various lengths. The coding sequences for collagen in this clone add up to approximately 1,800 bp, which correspond to about 1/3 of alpha 2 collagen mRNA. DNA sequence analysis of a small coding segment of lambda g COL 204 reveals a characteristic collagen type sequence which encodes for an amino acid sequence identical to a sequence found in calf alpha 2 collagen. The sequence of this region of the protein has not yet been determined for the chick alpha 2 collagen.

© 2021 EMBL | Privacy Policy