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  • Title: Directing transcription of an RNA polymerase III gene via GAL4 sites.
    Author: Marsolier MC, Chaussivert N, Lefebvre O, Conesa C, Werner M, Sentenac A.
    Journal: Proc Natl Acad Sci U S A; 1994 Dec 06; 91(25):11938-42. PubMed ID: 7991561.
    Abstract:
    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 (tau) or TFIIIB. Expression of chimeric tau 138 or tau 131 (but not tau 95) 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-(1-147)-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.
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