title: Proteasomal degradation of unassembled mutant type I collagen pro-alpha1(I) chains.
abstract: We have previously shown that type I procollagen pro-alpha1(I) chains from an osteogenesis imperfecta patient (OI26) with a frameshift mutation resulting in a truncated C-propeptide, have impaired assembly, and are degraded by an endoplasmic reticulum-associated pathway (Lamandé, S. R., Chessler, S. D., Golub, S. B., Byers, P. H., Chan, D., Cole, W. G., Sillence, D. O. and Bateman, J. F. (1995) J. Biol. Chem. 270, 8642-8649). To further explore the degradation of procollagen chains with mutant C-propeptides, mouse Mov13 cells, which produce no endogenous pro-alpha1(I), were stably transfected with a pro-alpha1(I) expression construct containing a frameshift mutation that predicts the synthesis of a protein 85 residues longer than normal. Despite high levels of mutant mRNA in transfected Mov13 cells, only minute amounts of mutant pro-alpha1(I) could be detected indicating that the majority of the mutant pro-alpha1(I) chains synthesized are targeted for rapid intracellular degradation. Degradation was not prevented by brefeldin A, monensin, or NH(4)Cl, agents that interfere with intracellular transport or lysosomal function. However, mutant pro-alpha1(I) chains in both transfected Mov13 cells and OI26 cells were protected from proteolysis by specific proteasome inhibitors. Together these data demonstrate for the first time that procollagen chains containing C-propeptide mutations that impair assembly are degraded by the cytoplasmic proteasome complex, and that the previously identified endoplasmic reticulum-associated degradation of mutant pro-alpha1(I) in OI26 is mediated by proteasomes.
authors.affiliation: Department of Paediatrics, Orthopaedic Molecular Biology Research Unit, University of Melbourne, Royal Children's Hospital, Parkville, Victoria 3052, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Fitzgerald
authors.forename: S R
authors.initials: SR
authors.lastname: Lamandé
authors.forename: J F
authors.initials: JF
authors.lastname: Bateman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D018388:Codon, Terminator; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D005091:Exons; D016368:Frameshift Mutation / Q000235:genetics*; D006801:Humans; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism*; D010013:Osteogenesis Imperfecta / Q000235:genetics; D011347:Procollagen / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012016:Reference Values; D016415:Sequence Alignment; D012867:Skin / Q000378:metabolism*; D014162:Transfection
chemical_list: D018388:Codon, Terminator; D009097:Multienzyme Complexes; D011347:Procollagen; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex