Cloning from a mouse osteoblastic cell line of a set of transforming-growth-factor-beta 1-regulated genes, one of which seems to encode a follistatin-related polypeptide.
Transforming growth factor(TGF)beta 1 is a potent inhibitor of growth in mouse osteoblastic MC3T3-E1 cells. To isolate genes that are induced by TGF beta 1, the differential screening method was adopted using a cDNA library constructed from cells treated with TGF beta 1 for 4 h. Six independent cDNA clones were isolated (TGF beta-stimulated clone, TSC-5, TSC-36, TSC-115, TSC-128, TSC-160 and TSC-161), the expression of which was increased by TGF beta 1-treatment with maximal expression at 6-10 h. The steady-state levels of TSC-36, TSC-128 and TSC-160 increased almost tenfold, whereas those of TSC-5, TSC-115 and TSC-161 were elevated at most threefold. From partial nucleotide sequences, TSC-160 was found to be identical to rrg (ras-recision gene, lysyl oxidase), and TSC-115 had 80% similarity with tropomyosin cDNA, whereas other genes seemed novel. Expression of TSC-36 and TSC-160 was dramatically decreased in v-Ki-ras-transformed MC3T3 cells or in transformed NIH 3T3 cells (DT), and was recovered to normal levels in a flat revertant (C11). A nearly full-length copy of TSC-36 cDNA was isolated, and an open reading frame indicated that it encodes a protein of 35 kDa. An antiserum was raised against the C-terminal peptide predicted from the nucleotide sequence, and a polypeptide with an approximate molecular mass of 38 kDa was detected in cultured medium of MC3T3-E1 cells. The amino acid sequence of TSC-36 protein was found to have some similarity with follistatin, an activin-binding protein, and a limited similarity with the secreted protein rich in cysteine (SPARC).
Shibanuma
M
M
Department of Microbiology, Showa University School of Pharmaceutical Sciences, Tokyo, Japan.
Mashimo
J
J
Mita
A
A
Kuroki
T
T
Nose
K
K
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D038681:Follistatin; D005786:Gene Expression Regulation; D011905:Genes, ras; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000737:chemistry* / Q000378:metabolism; D008249:Protein-Lysine 6-Oxidase / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D018076:DNA, Complementary; D038681:Follistatin; D006023:Glycoproteins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D008249:Protein-Lysine 6-Oxidase