Yeast RNA polymerase II transcription in vitro is inhibited in the presence of nucleotide excision repair: complementation of inhibition by Holo-TFIIH and requirement for RAD26.
The Saccharomyces cerevisiae transcription factor IIH (TFIIH) is essential both for transcription by RNA polymerase II (RNAP II) and for nucleotide excision repair (NER) of damaged DNA. We have established cell extracts which support RNAP II transcription from the yeast CYC1 promoter or NER of transcriptionally silent damaged DNA on independent plasmid templates and substrates. When plasmid templates and substrates for both processes are simultaneously incubated with these extracts, transcription is significantly inhibited. This inhibition is strictly dependent on active NER and can be complemented with purified holo-TFIIH. These results suggest that in the presence of active NER, TFIIH is preferentially mobilized from the basal transcription machinery for use in NER. Inhibition of transcription in the presence of active NER requires the RAD26 gene, the yeast homolog of the human Cockayne syndrome group B gene (CSB).
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Department of Pathology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Feaver
W J
WJ
Friedberg
E C
EC
D018797:Cell Cycle Proteins; D002474:Cell-Free System; D003574:Cytochrome c Group / Q000096:biosynthesis / Q000235:genetics; D045304:Cytochromes c; D004260:DNA Repair; D005656:Fungal Proteins / Q000378:metabolism*; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000378:metabolism*; D019473:Transcription Factors, TFII; D014158:Transcription, Genetic; D014466:Ultraviolet Rays / Q000009:adverse effects
C014059:CYC1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D003574:Cytochrome c Group; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; C490039:TAF6 protein, S cerevisiae; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D019473:Transcription Factors, TFII; C088086:rad26 protein, S pombe; D051758:Transcription Factor TFIIH; D045304:Cytochromes c; D012319:RNA Polymerase II