Abstract

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.

Highlights

  • The general transcription factor (TF)1 IID plays a critical role in transcription initiation of protein-coding genes by RNA polymerase II

  • The WD repeat is a conserved sequence motif usually ending with Trp-Asp (WD), and WD repeat-containing proteins are implicated in a wide variety of cellular functions [23]. yTAF90 and hTAF100 are components of histone acetylase complexes distinct from TFIID such as the Spt-Ada-gcn5ϩ and examined whether its product (Gcn5) acetyltransferase (SAGA) and TATA-binding protein (TBP)-free TAFIIcontaining (TFTC) complexes [24, 25]

  • We have identified two WD repeat-containing TBP-associated factors (TAFs) in fission yeast that may regulate genes involved in cell cycle progression

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Summary

TAF Genes as Multicopy Suppressors of Mitotic Arrest

We show that both TAF72 and TAF73 are associated with TBP and other TAFs, whereas only TAF72 is associated with Gcn, a putative histone acetylase. We show that taf72ϩ and taf73ϩ suppress a mutation in the cut9ϩ gene. These results suggest that TAF72 and TAF73 may regulate the expression of genes involved in progression through the M phase of the cell cycle

EXPERIMENTAL PROCEDURES
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