UPSTREAM STIMULATORY FACTOR PROTEINS ARE MAJOR COMPONENTS OF THE GLUCOSE RESPONSE COMPLEX OF THE L-TYPE PYRUVATE-KINASE GENE PROMOTER

L-type pyruvate kinase (L-PK) gene transcription is induced by glucose through its glucose response element (GIRE) composed of two degenerated E boxes able to bind in vitro ubiquitous upstream stimulator factor (USF) proteins. Here we demonstrate in vivo, by transient transfections in hepatoma cells, that (i) native USF proteins synthesized from expression vectors can act as transactivators of the L-PK promoter via the GIRE, stimulating transcription without glucose and, therefore, decreasing the glucose responsiveness of the promoter; (ii) expression of the truncated USF proteins, able to bind the GIRE but devoid of the NH2-terminal activation domain, represses the activation of the L-PK promoter by glucose; and (iii) a similar repression of the glucose effect is observed upon expression of mutant USF proteins devoid of the basic DNA binding domain, able to dimerize with endogenous USF but not to bind the GIRE. We conclude that USF proteins are components of the transcriptional glucose response complex assembled on the L-PK gene promoter.