The multiple roles of transcription/repair factor TFIIH

Svejstrup, Jesper Q.; Vichi, Paul; Egly, Jean‐Marc

doi:10.1016/s0968-0004(96)10046-3

Cited by 171 publications

(118 citation statements)

References 41 publications

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“…In S. cerevisiae and higher eukaryotic cells, TFIIH contains nine subunits (25). To purify S. pombe TFIIH, we introduced a TAP-tag on the C terminus of Pmh1.…”

Section: Resultsmentioning

confidence: 99%

Mediator Influences Schizosaccharomyces pombe RNA Polymerase II-dependent Transcription in Vitro

Spåhr

Khorosjutina

Baraznenok

et al. 2003

Journal of Biological Chemistry

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The fission yeast Schizosaccharomyces pombe has proved an important model system for cross-species comparative studies of many fundamental processes in the eukaryotic cell, such as cell cycle control and DNA replication. The RNA polymerase II transcription machinery is, however, still relatively poorly understood in S. pombe, partially due to the absence of a reconstituted in vitro transcription system. We have now purified S. pombe RNA polymerase II and its general initiation factors TFIIB, TFIIF, TFIIE, and TFIIH to near homogeneity. These factors enable RNA polymerase II to initiate transcription from the S. pombe alcohol dehydrogenase promoter (adh1p) when combined with Saccharomyces cerevisiae TATA-binding protein. We use our reconstituted system to examine effects of Mediator on basal transcription in vitro. S. pombe Mediator exists in two distinct forms, a free form, which contains the spSrb8, spTrap240, spSrb10, and spSrb11 subunits, and a smaller form, which lacks these four subunits and associates with RNA polymerase II to form a holoenzyme. We find that spSrb8/spTrap240/spSrb10/spSrb11 containing Mediator repress basal transcription, whereas Mediator lacking these subunits has a stimulatory effect on transcription. Our findings thus demonstrate that the spSrb8/spTrap240/spSrb10/spSrb11 subcomplex governs the ability of Mediator to stimulate or repress basal transcription in vitro.

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“…In S. cerevisiae and higher eukaryotic cells, TFIIH contains nine subunits (25). To purify S. pombe TFIIH, we introduced a TAP-tag on the C terminus of Pmh1.…”

Section: Resultsmentioning

confidence: 99%

Mediator Influences Schizosaccharomyces pombe RNA Polymerase II-dependent Transcription in Vitro

Spåhr

Khorosjutina

Baraznenok

et al. 2003

Journal of Biological Chemistry

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“…The cyclin-dependent kinase CDK7, present in the multisubunit TFIIH complex has been widely postulated to function as a major CTD kinase in vivo (for review, see Akoulitchev et al 1995;Svejstrup et al 1996), leading several groups to examine the possibility that Tat may target TFIIH directly (Fig. 2).…”

Section: Interactions Between Tat and The Ctd Kinase Cak (Tfiih)mentioning

confidence: 99%

Taking a new TAK on Tat transactivation

1997

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“…The unphosphorylated form of Pol II (Pol IIa) is preferentially recruited into a PIC reconstituted with purified general transcription factors (30). CTD phosphorylation may initially occur between transcription initiation and the transition from initiation to elongation, converting Pol IIa to the phosphorylated form (Pol IIo) (34,35,59). Recently, it has been demonstrated that CTD phosphorylation is also important for recruiting the mRNA processing enzymes to the nascent transcript, presumably reflecting the fact that mRNA processing (splicing, capping, and polyadenylation) occurs during and/or after transcription (6,33).…”

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confidence: 99%

Studies of Nematode TFIIE Function Reveal a Link between Ser-5 Phosphorylation of RNA Polymerase II and the Transition from Transcription Initiation to Elongation

Yamamoto

Watanabe

Spek

et al. 2001

Molecular and Cellular Biology

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The general transcription factor TFIIE plays important roles in transcription initiation and in the transition to elongation. However, little is known about its function during these steps. Here we demonstrate for the first time that TFIIH-mediated phosphorylation of RNA polymerase II (Pol II) is essential for the transition to elongation. This phosphorylation occurs at serine position 5 (Ser-5) of the carboxy-terminal domain (CTD) heptapeptide sequence of the largest subunit of Pol II. In a human in vitro transcription system with a supercoiled template, this process was studied using a human TFIIE (hTFIIE) homolog from Caenorhabditis elegans (ceTFIIE␣ and ceTFIIE␤). ceTFIIE␤ could partially replace hTFIIE␤, whereas ceTFIIE␣ could not replace hTFIIE␣. We present the studies of TFIIE binding to general transcription factors and the effects of subunit substitution on CTD phosphorylation. As a result, ceTFIIE␣ did not bind tightly to hTFIIE␤, and ceTFIIE␤ showed a similar profile for binding to its human counterpart and supported an intermediate level of CTD phosphorylation. Using antibodies against phosphorylated serine at either Ser-2 or Ser-5 of the CTD, we found that ceTFIIE␤ induced Ser-5 phosphorylation very little but induced Ser-2 phosphorylation normally, in contrast to wild-type hTFIIE, which induced phosphorylation at both Ser-2 and Ser-5. In transcription transition assays using a linear template, ceTFIIE␤ was markedly defective in its ability to support the transition to elongation. These observations provide evidence of TFIIE involvement in the transition and suggest that Ser-5 phosphorylation is essential for Pol II to be in the processive elongation form.In eukaryotes, transcription of protein-encoding genes by RNA polymerase II (Pol II) is the first step in expression of those genes (for reviews, see references 4, 35, 44, and 51). Two sequential stages are now recognized in the establishment of Pol II processivity: transcription initiation and the transition from initiation to elongation. At initiation, five general transcription factors (TFIIB, TFIID, TFIIE, TFIIF, and TFIIH) together with Pol II form the preinitiation complex (PIC) on the core promoter. Two models of PIC formation have been proposed on the basis of recent analyses. One model involves stepwise association of the general transcription factors and Pol II on promoter DNA, while the other model entails promoter sequences binding to a preassembled Pol II holoenzyme that contains most of the general transcription factors as well as SRB (suppressor of RNA polymerase B)-and Med-containing complex (reviewed in references 3 and 22). In vitro analyses of stepwise assembly of the PIC using purified factors have demonstrated that TFIIE joins the complex at a position near the transcription start site (between positions Ϫ14 and Ϫ2), after Pol II and TFIIF have joined the complex (25, 49). TFIIE then recruits TFIIH, and these two factors stabilize and activate the PIC, resulting in isomerization of double-stranded (ds) promoter DNA (promoter me...

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The multiple roles of transcription/repair factor TFIIH

Cited by 171 publications

References 41 publications

Mediator Influences Schizosaccharomyces pombe RNA Polymerase II-dependent Transcription in Vitro

Mediator Influences Schizosaccharomyces pombe RNA Polymerase II-dependent Transcription in Vitro

Taking a new TAK on Tat transactivation

Studies of Nematode TFIIE Function Reveal a Link between Ser-5 Phosphorylation of RNA Polymerase II and the Transition from Transcription Initiation to Elongation

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