Main content area

Directing transcription of an RNA polymerase III gene via GAL4 sites

Marsolier, M.C., Chaussivert, N., Lefebvre, O., Conesa, C., Werner, M., Sentenac, A.
Proceedings of the National Academy of Sciences of the United States of America 1994 v.91 no.25 pp. 11938-11942
Saccharomyces cerevisiae, structural genes, DNA-directed RNA polymerase, recombinant DNA, nucleotide sequences, DNA-binding proteins, transcription (genetics), transcription factors, binding sites
A yeast chimeric RNA polymerase III transcription system was constructed to explore the ordered, multistep process of gene activation in vivo. A promoter-deficient U6 RNA gene harboring GAL4-binding sites could be reactivated by fusing the GAL4 DNA-binding domain to components of the general transcription factor TFIIIC (T) or TFIIIB. Expression of chimeric T138 or T131 (but not T95) subunits activated transcription from GAL4-binding sites located at various positions, including upstream of or within the gene. The function(s) of the B block binding domain of TFIIIC was provided by the fused GAL4-(1-147) domain. The GAL4-(1147)TFIIIB70 fusion protein acted at a distance like an activator of transcription. In contrast, none of the 10 different GAL4-(1-147)-polymerase subunit fusions was able to induce transcription, suggesting that RNA polymerase recruitment is not sufficient to initiate transcription.