Mitsuzawa H, Seino H, Yamao F, Ishihama A
Divisions of Molecular Genetics and Mutagenesis, Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
J Biol Chem. 2001 May 18;276(20):17117-24. doi: 10.1074/jbc.M100248200. Epub 2001 Mar 9.
The general transcription factor IID consists of the TATA-binding protein (TBP) and multiple TBP-associated factors (TAFs). Here we report the isolation of two related TAF genes from the fission yeast Schizosaccharomyces pombe as multicopy suppressors of a temperature-sensitive mutation in the ubiquitin-conjugating enzyme gene ubcP4(+). The ubcP4(ts) mutation causes cell cycle arrest in mitosis, probably due to defects in ubiquitination mediated by the anaphase-promoting complex/cyclosome. One multicopy suppressor is the previously reported gene taf72(+), whereas the other is a previously unidentified gene named taf73(+). We show that the taf73(+) gene, like taf72(+), is essential for cell viability. The taf72(+) and taf73(+) genes encode proteins homologous to WD repeat-containing TAFs such as human TAF100, Drosophila TAF80/85, and Saccharomyces cerevisiae TAF90. We demonstrate that TAF72 and TAF73 proteins are present in the same complex with TBP and other TAFs and that TAF72, but not TAF73, is associated with the putative histone acetylase Gcn5. We also show that overexpression of TAF72 or TAF73 suppresses the cell cycle arrest in mitosis caused by a mutation in the anaphase-promoting complex/cyclosome subunit gene cut9(+). These results suggest that TAF72 and TAF73 may regulate the expression of genes involved in ubiquitin-dependent proteolysis during mitosis. Our study thus provides evidence for a possible role of WD repeat-containing TAFs in the expression of genes involved in progression through the M phase of the cell cycle.
通用转录因子IID由TATA结合蛋白(TBP)和多个TBP相关因子(TAF)组成。在此,我们报告从裂殖酵母粟酒裂殖酵母中分离出两个相关的TAF基因,作为泛素结合酶基因ubcP4(+)中温度敏感突变的多拷贝抑制子。ubcP4(ts)突变导致细胞周期在有丝分裂期停滞,这可能是由于后期促进复合物/细胞周期体介导的泛素化缺陷所致。一个多拷贝抑制子是先前报道的基因taf72(+),而另一个是先前未鉴定的基因,命名为taf73(+)。我们表明taf73(+)基因与taf72(+)一样,对细胞活力至关重要。taf72(+)和taf73(+)基因编码与含WD重复序列的TAF同源的蛋白质,如人TAF100、果蝇TAF80/85和酿酒酵母TAF90。我们证明TAF72和TAF73蛋白与TBP和其他TAF存在于同一复合物中,并且TAF72而非TAF73与推定的组蛋白乙酰转移酶Gcn5相关。我们还表明,TAF72或TAF73的过表达可抑制由后期促进复合物/细胞周期体亚基基因cut9(+)突变引起的有丝分裂期细胞周期停滞。这些结果表明,TAF72和TAF73可能在有丝分裂期间调节参与泛素依赖性蛋白水解的基因的表达。因此,我们的研究为含WD重复序列的TAF在参与细胞周期M期进程的基因表达中可能发挥的作用提供了证据。