title: Partial characterization of a low molecular weight human collagen that undergoes alternative splicing.
abstract: A cDNA library prepared from RNA isolated from a cultured human tumor cell line, HT-1080, was screened with a mouse cDNA clone coding for part of the -Gly-Xaa-Yaa- domain of the alpha 2(IV) collagen chain. Four overlapping cDNA clones were characterized that coded for a low molecular weight human collagen. The cDNA clones did not, however, code for the short-chain collagens, types IX and X. The amino acid sequences derived from the clones resembled type IV collagen in that there were short interruptions in the repeating -Gly-Xaa-Yaa- sequence. The noncollagenous, carboxyl-terminal domain was, however, much shorter and contained only 18 amino acid residues. Interestingly, one of the cDNA clones contained an additional 36 nucleotides not found in an overlapping clone. The 36 nucleotides encoded four -Gly-Xaa-Yaa- repeats without changing the reading frame. Nuclease S1 mapping demonstrated that the difference between the clones was due to existence of two different mRNAs. A synthetic 24-residue peptide corresponding to the last two -Gly-Xaa-Yaa- triplets and the entire carboxyl-terminal domain was used to generate polyclonal antibodies. Electrophoretic transfer blot analysis of HT-1080 cells and normal human skin fibroblasts identified two polypeptides, Mr 67,000 and Mr 62,000, that were sensitive to bacterial collagenase.
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003094:Collagen / Q000235:genetics*; D004270:DNA, Circular / Q000235:genetics; D004274:DNA, Recombinant; D004586:Electrophoresis / Q000379:methods; D005815:Genetic Code; D005821:Genetic Techniques; D006801:Humans; D008970:Molecular Weight; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D003062:Codon; D004270:DNA, Circular; D004274:DNA, Recombinant; D012333:RNA, Messenger; D003094:Collagen