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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.