Transcription inhibition of the somatic-type phosphoglycerate kinase 1 gene in vitro by a testis-specific factor that recognizes a sequence similar to the binding site for Ets oncoproteins.
To elucidate the mechanism by which transcription of the somatic-type phosphoglycerate kinase 1 gene is inactivated during mammalian spermatogenesis, we examined the presence of specific transcription inhibitor(s) in the testis by a cell-free transcription system. Transcription of the mouse phosphoglycerate kinase 1 gene using nuclear extracts of the rat liver was significantly inhibited by the addition of testis extracts, whereas brain extracts had little effect. Transcription inhibition required the binding of a testis-specific factor, designated TIN-1, to the region between positions -268 and -259 relative to transcription initiation site at +1. This region had the sequence 5'-AGGAAGTTCC-3' that includes an inverted repeat of the binding motif, 5'-GGAA-3', for the oncoprotein Ets. A UV-crosslinking experiment revealed that 43- and 45-kDa polypeptides present in testis extracts bind to that sequence. These results suggest that a testis-specific transcription inhibitor TIN-1 inactivates the phosphoglycerate kinase 1 gene in the mammalian spermatogenic pathway.
Goto
M
M
Faculty of Pharmaceutical Sciences, Kanazawa University, Ishikawa, Japan.
Tamura
T
T
Mikoshiba
K
K
Masamune
Y
Y
Nakanishi
Y
Y
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002457:Cell Extracts; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D008297:Male; D008969:Molecular Sequence Data; D010735:Phosphoglycerate Kinase / Q000235:genetics*; D051381:Rats; D015682:Retroviridae Proteins, Oncogenic / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics; D013696:Temperature; D013737:Testis / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic
D002457:Cell Extracts; D004268:DNA-Binding Proteins; D015682:Retroviridae Proteins, Oncogenic; C050379:oncogene proteins v-ets; D004247:DNA; D010735:Phosphoglycerate Kinase