Identification of a yeast protein homologous in function to the mammalian general transcription factor, TFIIA.
The yeast homolog of the mammalian RNA polymerase II general transcription factor TFIIA has been identified by complementation of a mammalian in vitro transcription system depleted for TFIIA. Like the mammalian factor, the yeast protein does not bind DNA, alters the size of the TFIID DNase I footprint at the adenovirus major late promoter, and forms specific TFIIA-TFIID-DNA complexes which are stable during electrophoresis in native acrylamide gels. The partially purified yeast factor was used to investigate its effect on the binding of TFIID to the major late promoter. Contrary to earlier models, we find that TFIIA does not significantly change the affinity or kinetics of TFIID binding, suggesting that it acts by altering the conformation of TFIID and/or by serving as a bridge between TFIID and the other general transcription factors.
Hahn
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S
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Buratowski
S
S
Sharp
P A
PA
Guarente
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L
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D007700:Kinetics; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D035362:Transcription Factor TFIID; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
D004279:DNA, Viral; D005656:Fungal Proteins; D035362:Transcription Factor TFIID; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors; D003850:Deoxyribonuclease I