A silencer-like cis element for the testis-specific phosphoglycerate-kinase-2-encoding gene

Abstract

Phosphoglycerate kinase (PGK), a glycolytic enzyme, possesses two isozymes: somatic-type PGK-1 and testis-specific PGK-2, encoded by distinct genes. Tissue-specific expression of the two PGK-encoding genes (Pgk) seems to be transcriptionally controlled, since tissue distribution of the mRNAs coincides well with that of the proteins. In the present study, we determined the cis-acting DNA elements that regulate the transcription of mouse Pgk-2. A transient expression assay of DNAs having various portions of the Pgk-2 upstream region linked to the chloramphenicol acetyltransferase (CAT)-encoding gene (cat) was performed using mouse cell lines that exclusively express Pgk-1. A substantial increase in cat expression was observed when the region between nucleotides (nt) -1404 and -685, relative to the most distal transcription start point at nt +1, was lost. This cis-acting region appeared to function as a silencer, since it repressed cat expression independently of either orientation to or distance from the Pgk-2 promoter. Moreover, the cis element inhibited Pgk-2 transcription with no effect on Pgk-1 transcription in a cell-free system using nuclear extracts of rat liver. These results suggest that a silencer-like negative cis element is responsible, at least partly, for tissue-specific transcription of Pgk-2.