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Molecular cloning of a mouse 47‐kDa heat‐shock protein (HSP47), a collagen‐binding stress protein, and its expression during the differentiation of F9 teratocarcinoma cells

TAKECHI, Hajime, HIRAYOSHI, Kazunori, NAKAI, Akira, KUDO, Hitoshi, SAGA, Shinsuke, NAGATA, Kazuhiro
European journal of biochemistry 1992 v.206 no.2 pp. 323-329
cDNA libraries, chicks, collagen, complementary DNA, endoplasmic reticulum, gene expression, heat, heat shock proteins, humans, messenger RNA, mice, molecular cloning, precipitin tests, proteinase inhibitors, retinoic acid, serine proteinases, signal peptide, two-dimensional gel electrophoresis
A 47‐kDa heat‐shock protein (HSP47) is a major collagen‐binding stress protein residing in the endoplasmic reticulum, and is assumed to be a molecular chaperone specific to collagen. Two‐dimensional gel electrophoresis and immunoprecipitation studies showed that the expression of HSP47 was significantly induced during the differentiation of mouse teratocarcinoma F9 cells by treatment with retinoic acid alone or with retinoic acid and dibutyryladenosine 3′,5′‐phosphate. The induction of type‐IV collagen was also observed during F9‐cell differentiation. For further analysis, we cloned cDNA encoding mouse HSP47 from a cDNA library of BALB/c 3T3 cells and performed Northern‐blot analysis. The cDNA contained a signal sequence at the N‐terminus and an endoplasmic‐reticulum‐retention signal, RDEL, at the C‐terminus. An homology search revealed that mouse HSP47, as well as chick HSP47, belonged to the serine protease inhibitor superfamily. While chick HSP47 mRNA was 4.5 kb with a long (2‐kb) 3′ untranslated region, mouse and human HSP47 mRNA were 2.5 kb, with a 0.8‐kb 3′ untranslated region. Northern‐blot analysis revealed that the concurrent induction of HSP47 and type‐IV collagen during F9‐cell differentiation, and the transient induction of HSP47 after heat shock was regulated at the level of mRNA accumulation. These results suggested that HSP47 was closely related to collagens in terms of its expression as well as in its functional relevance.