Analysis of selective gene activation in yeast by differential display.
TATA box binding protein-associated factors (TAF(II)s) are dispensable for transcription of most genes in yeast. To further investigate the in vivo functions of TAF(II)s, differential display was used to identify a small subset of yeast genes whose transcription is dependent on yeast TAF(II)145, the core TAF(II) component that contacts TATA box-binding protein (TBP). Messenger RNA profiles derived from a wild-type TAF(II)145 strain and a temperature-sensitive taf(II)1145 strain were analyzed. Those genes whose messenger RNA level was greatly reduced in the temperature-sensitive taf(II)145 strain were cloned and further characterized. The procedure for performing differential display described here is modified from protocols provided by the manufacturer (Display Systems) and optimized for the yeast system.
Howard Hughes Medical Institute, Program in Molecular Medicine, University of Massachusetts Medical Center, Worcester 01605, USA.
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D001665:Binding Sites / Q000235:genetics; D015152:Blotting, Northern; D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation; D015003:Yeasts / Q000235:genetics*
D017931:DNA Primers; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C106785:TAF1 protein, S cerevisiae; D035381:TATA-Binding Protein Associated Factors; D035181:TATA-Box Binding Protein; D035362:Transcription Factor TFIID; D014157:Transcription Factors