Introduction of the human pro alpha 1(I) collagen gene into pro alpha 1(I)-deficient Mov-13 mouse cells leads to formation of functional mouse-human hybrid type I collagen.
The Mov-13 mouse strain carries a retroviral insertion in the pro alpha 1(I) collagen gene that prevents transcription of the gene. Cell lines derived from homozygous embryos do not express type I collagen although normal amounts of pro alpha 2 mRNA are synthesized. We have introduced genomic clones of either the human or mouse pro alpha 1(I) collagen gene into homozygous cell lines to assess whether the human or mouse pro alpha 1(I) chains can associate with the endogenous mouse pro alpha 2(I) chain to form stable type I collagen. The human gene under control of the simian virus 40 promoter was efficiently transcribed in the transfected cells. Protein analyses revealed that stable heterotrimers consisting of two human alpha 1 chains and one mouse alpha 2 chain were formed and that type I collagen was secreted by the transfected cells at normal rates. However, the electrophoretic migration of both alpha 1(I) and alpha 2(I) chains in the human-mouse hybrid molecules were retarded, compared to the alpha (I) chains in control mouse cells. Inhibition of the posttranslational hydroxylation of lysine and proline resulted in comigration of human and mouse alpha 1 and alpha 2 chains, suggesting that increased posttranslational modification caused the altered electrophoretic migration in the human-mouse hybrid molecules. Amino acid sequence differences between the mouse and human alpha chains may interfere with the normal rate of helix formation and increase the degree of posttranslational modifications similar to those observed in patients with lethal perinatal osteogenesis imperfecta. The Mov-13 mouse system should allow us to study the effect specific mutations introduced in transfected pro alpha 1(I) genes have on the synthesis, assembly, and function of collagen I.
Schnieke
A
A
Dziadek
M
M
Bateman
J
J
Mascara
T
T
Harbers
K
K
Gelinas
R
R
Jaenisch
R
R
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D004622:Embryo, Mammalian; D005796:Genes; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D051379:Mice; D011347:Procollagen / Q000096:biosynthesis / Q000235:genetics*; D055503:Protein Multimerization
C000716128:Col1a1 protein, mouse; D000090602:Collagen Type I, alpha 1 Chain; D011347:Procollagen; D003094:Collagen