The silencer of mouse Pgk-2 consists of two discrete DNA elements that individually have no effect

Pgk-2 encodes a testis-specific phosphoglycerate kinase isozyme (PGK), and the expression of mouse Pgk-2 is activated during the spermatogenic pathway at the pachytene spermatocyte stage. We previously reported the identification of a silencer-like cis-acting element in a region between nucleotides (nt) -1404 and -685 of mouse Pgk-2, which could be responsible for the repression of Pgk-2 expression in somatic tissues and pre-meiotic testicular germ cells. In the present study, the silencer was precisely located within an 87-bp region between nt -882 and -796. This region contained two distinct sequences that individually bound factors present in nuclear extracts of mouse cultured cells and rat tissues. The two sequences, when aligned in tandem upstream from the Pgk-2 promoter, inhibited cat expression driven by the promoter in mouse erythroleukemia cells, whereas either sequence alone did not show any effect. The results indicate that the Pgk-2 silencer consists of two distinct DNA elements which do not individually influence promoter activity. Binding of distinct nuclear factors to each DNA element appeared to be required for the silencer action.