TAFII250-dependent transcription of cyclin A is directed by ATF activator proteins.
A specific mutation in TAFII250, the largest subunit of the transcription factor TFIID, disrupts cell growth control in the temperature-sensitive mutant hamster cell line ts13. Transcription from the cyclin A and D1 but not the c-fos and myc promoters is also dramatically reduced in ts13 cells at the nonpermissive temperature. These findings provide an intriguing link between TAF-mediated transcriptional regulation and cell cycle progression. Here we report the mapping of an enhancer element in the cyclin A promoter (TSRE) that responds to mutations in TAFII250. An analysis of chimeric promoter constructs reveals that the cyclin A TSRE can confer TAFII250 dependence to the core promoter of c-fos. In addition, reciprocal hybrid promoter constructs suggest that TAFII250 also contributes to the transcriptional properties of the cyclin A core promoter. We have purified and identified cellular activators that specifically bind to the TSRE and mediate transcription in a TAFII250-dependent manner. By micropeptide sequencing, we determined that TSRE-binding proteins include members of the activating transcription factor (ATF) family. These results suggest that the ts13 mutation of TAFII250 has compromised the ability of TFIID to mediate activation of transcription by specific enhancer factors such as ATF, as well as to perform certain core promoter functions. These defects in TAFII250 apparently result in the down-regulation of key molecules, such as cyclin A, which may be responsible for the ts13 cell cycle arrest phenotype.
Wang
E H
EH
Department of Pharmacology, School of Medicine, University of Washington, Seattle, Washington 98195-7280 USA. ehwang@u.washington.edu
Zou
S
S
Tjian
R
R
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D019925:Cyclin A / Q000096:biosynthesis* / Q000235:genetics; D019938:Cyclin D1 / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D016762:Genes, fos; D016259:Genes, myc; D006367:HeLa Cells; D051548:Histone Acetyltransferases; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D035381:TATA-Binding Protein Associated Factors; D013696:Temperature; D015534:Trans-Activators / Q000378:metabolism*; D035362:Transcription Factor TFIID; D014158:Transcription, Genetic; D014162:Transfection
D019925:Cyclin A; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D035381:TATA-Binding Protein Associated Factors; D015534:Trans-Activators; D035362:Transcription Factor TFIID; D019938:Cyclin D1; D008156:Luciferases; D051548:Histone Acetyltransferases; C069001:TATA-binding protein associated factor 250 kDa