Cloning of TPA-inducible early (TIE) genes by differential hybridization using TPA-nonresponsive variant of mouse 3T3-L1 cells.
The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) induces DNA synthesis in quiescent 3T3-L1 cells but not in its variant VT-1 cells. A lambda gt10 cDNA library was constructed using poly(A)+ RNA from 3T3-L1 cells that were stimulated by TPA for 20 min. Radioactive cDNA probes were prepared from mRNAs of TPA-treated 3T3-L1 and VT-1 cells and used for screening of the 3T3-L1 cDNA library by differential hybridization. Nine of 6000 phase plaques hybridized only to the 3T3-L1 cDNA probe. Analysis of the nucleotide sequence of five of these clones indicated a high degree of homology with human or mouse type I and type III collagen genes. Three other independent clones showed no homology with any known DNA sequences. These isolated clones of TPA-inducible early (TIE) genes may be useful to study the signal transduction pathway of phorbol esters.
Amagai
M
M
Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.
Inokuchi
Y
Y
Nishikawa
T
T
Shimizu
Y
Y
Shimizu
N
N
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics; D015342:DNA Probes; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D015342:DNA Probes; D012333:RNA, Messenger; D003094:Collagen; D013755:Tetradecanoylphorbol Acetate