Multiple forms of chicken alpha 3(VI) collagen chain generated by alternative splicing in type A repeated domains

Type VI collagen is a structurally unique component widely distributed in connective tissues. Its molecular structure consists of monomers that have the potential to assemble intracellularly into dimers and tetramers which, once secreted, can form microfilaments by end-to-end association. Individual monomers are composed of chains of Mr = approximately 140,000 (alpha 1 and alpha 2) and greater than 300,000 (alpha 3). Type VI collagen molecules contain a short triple helix with large globular domains at both ends. These domains are made for their greatest part of repetitive units similar to type A repeats of von Willebrand Factor. The alpha 3(VI) chain, contributing most of the mass of the NH2-terminal globule, appeared heterogenous both at the mRNA and protein level. Several alpha 3(VI)-specific clones that lack the sequences corresponding to repeats A8 and A6 were isolated from a chicken aorta cDNA library. Northern blot hybridization of poly (A+)- enriched RNA from chicken gizzard with cDNA fragments corresponding to several individual type A repeats showed that A8- and A6-specific probes did not hybridize to the lower Mr transcripts. Clones spanning approximately 20 kb of the 5'-end of the alpha 3(VI) gene were isolated from a chicken genomic library and subjected to analysis by restriction mapping, Southern blotting, and selective sequencing of the intron-exon boundaries. At the most 5'-end of the gene an additional type A repeat (A9), previously undetected in cDNA clones, was identified. Furthermore, it was determined that the presumed signal peptide and repeats A9 through A6 are encoded within individual exons. Reverse transcription and polymerase chain reaction of aorta RNA suggested that a mechanism of alternative mRNA splicing by a phenomenon of exon skipping generates alpha 3(VI) isoform variants that contain different numbers of type A repeats. Immunohistochemistry of frozen sections of chicken embryo tissues with repeat-specific mAbs showed that an antibody directed against a conditional exon has a more restricted tissue distribution compared to an antibody against a constitutive exon.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  F. Bucciotti,et al.  Structural and functional features of the alpha 3 chain indicate a bridging role for chicken collagen VI in connective tissues. , 1990, Biochemistry.

[3]  P. Bonaldo,et al.  The carboxyl terminus of the chicken alpha 3 chain of collagen VI is a unique mosaic structure with glycoprotein Ib-like, fibronectin type III, and Kunitz modules. , 1989, The Journal of biological chemistry.

[4]  B. Olsen,et al.  Tissue-specific forms of type IX collagen-proteoglycan arise from the use of two widely separated promoters. , 1989, The Journal of biological chemistry.

[5]  Harold E. Dvorak,et al.  Reappearance of an embryonic pattern of fibronectin splicing during wound healing in the adult rat , 1989, The Journal of cell biology.

[6]  A. Colombatti,et al.  Type VI collagen: high yields of a molecule with multiple forms of alpha 3 chain from avian and human tissues. , 1989, Matrix.

[7]  R. Hynes,et al.  Alternative splicing of fibronectin is temporally and spatially regulated in the chicken embryo. , 1989, Development.

[8]  F. Bucciotti,et al.  Alpha 1 chain of chick type VI collagen. The complete cDNA sequence reveals a hybrid molecule made of one short collagen and three von Willebrand factor type A-like domains. , 1989 .

[9]  E. Koller,et al.  The globular domains of type VI collagen are related to the collagen‐binding domains of cartilage matrix protein and von Willebrand factor. , 1989, The EMBO journal.

[10]  F. Baralle,et al.  Cell type specific trans‐acting factors are involved in alternative splicing of human fibronectin pre‐mRNA. , 1989, The EMBO journal.

[11]  L. Zardi,et al.  A tumor-associated fibronectin isoform generated by alternative splicing of messenger RNA precursors , 1989, The Journal of cell biology.

[12]  G. Edelman,et al.  A detailed structural model of cytotactin: protein homologies, alternative RNA splicing, and binding regions. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Gulcher,et al.  An alternatively spliced region of the human hexabrachion contains a repeat of potential N-glycosylation sites. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Hynes,et al.  Patterns of fibronectin gene expression and splicing during cell migration in chicken embryos. , 1988, Development.

[15]  R. Glanville,et al.  Ultrastructure of type VI collagen in human skin and cartilage suggests an anchoring function for this filamentous network , 1988, The Journal of cell biology.

[16]  T. Pihlajaniemi,et al.  Gene structure for the alpha 1 chain of a human short-chain collagen (type XIII) with alternatively spliced transcripts and translation termination codon at the 5' end of the last exon. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[17]  M J Banda,et al.  Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping. , 1988, Science.

[18]  P. Bonaldo,et al.  Monoclonal antibodies for the different chains of chick type VI collagen. , 1988, Collagen and related research.

[19]  K. Doege,et al.  Alternative splicing generates two different mRNA species for rat link protein. , 1988, The Journal of biological chemistry.

[20]  F. Bucciotti,et al.  Isolation of cDNA clones corresponding to the Mr = 150,000 subunit of chick type VI collagen. , 1987, Biochemical and biophysical research communications.

[21]  D. Eyre,et al.  Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein. , 1987, The Biochemical journal.

[22]  P. Bonaldo,et al.  Biosynthesis of chick type VI collagen. II. Processing and secretion in fibroblasts and smooth muscle cells. , 1987, The Journal of biological chemistry.

[23]  P. Bonaldo,et al.  Biosynthesis of chick type VI collagen. I. Intracellular assembly and molecular structure. , 1987, The Journal of biological chemistry.

[24]  A. Kornblihtt,et al.  Identification of a third region of cell-specific alternative splicing in human fibronectin mRNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Timpl,et al.  Characterization of three constituent chains of collagen type VI by peptide sequences and cDNA clones. , 1987, European journal of biochemistry.

[26]  R. Hynes,et al.  Multiple sites of alternative splicing of the rat fibronectin gene transcript. , 1987, The EMBO journal.

[27]  L Peltonen,et al.  Alternative splicing of human elastin mRNA indicated by sequence analysis of cloned genomic and complementary DNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[28]  F. Baralle,et al.  Transformed human cells produce a new fibronectin isoform by preferential alternative splicing of a previously unobserved exon. , 1987, The EMBO journal.

[29]  C. Richardson,et al.  DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[30]  P Argos,et al.  Repeating structure of chick tropoelastin revealed by complementary DNA cloning. , 1987, Biochemistry.

[31]  R. Timpl,et al.  Type VI collagen in extracellular, 100-nm periodic filaments and fibrils: identification by immunoelectron microscopy , 1986, The Journal of cell biology.

[32]  G. Heijne.  A new method for predicting signal sequence cleavage sites. , 1986 .

[33]  K. Titani,et al.  cDNA sequences for human von Willebrand factor reveal five types of repeated domains and five possible protein sequence polymorphisms. , 1986, Biochemistry.

[34]  E. Engvall,et al.  Molecular assembly, secretion, and matrix deposition of type VI collagen , 1986, The Journal of cell biology.

[35]  R. Bruns,et al.  Beaded filaments and long-spacing fibrils: relation to type VI collagen. , 1984, Journal of ultrastructure research.

[36]  R. Timpl,et al.  Immunochemistry, genuine size and tissue localization of collagen VI. , 1984, European journal of biochemistry.

[37]  D. Troyer,et al.  A collagen-like glycoprotein of the extracellular matrix is the undegraded form of type VI collagen. , 1984, Biochemistry.

[38]  A. Kornblihtt,et al.  Human fibronectin: cell specific alternative mRNA splicing generates polypeptide chains differing in the number of internal repeats. , 1984, Nucleic acids research.

[39]  G. Hong,et al.  Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[40]  R. Timpl,et al.  Electron-microscopical approach to a structural model of intima collagen. , 1983, The Biochemical journal.

[41]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[42]  R. Burgeson.  New collagens, new concepts. , 1988, Annual review of cell biology.

[43]  B. Nadal-Ginard,et al.  Generation of protein isoform diversity by alternative splicing: mechanistic and biological implications. , 1987, Annual review of cell biology.

[44]  P. Sharp,et al.  Splicing of messenger RNA precursors. , 1987, Science.

[45]  G. von Heijne,et al.  A new method for predkting signal sequence cleavage sites , 2022 .

[46]  P. Sharp,et al.  Splicing of messenger RNA precursors. , 1985, Harvey lectures.

[47]  J. Messing.  New M13 vectors for cloning. , 1983, Methods in enzymology.

[48]  J. Messing.  [2] New M13 vectors for cloning , 1983 .