Specific interaction between the nonphosphorylated form of RNA polymerase II and the TATA-binding protein

Usheva, Anny; Maldonado, Edio; Goldring, Anat; Lu, Hua; Houbavi, Christo; Reinberg, Danny; Aloni, Yosef

doi:10.1016/0092-8674(92)90297-p

Cited by 215 publications

(140 citation statements)

References 42 publications

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“…It is now widely recognized that the CTD of Pol II is hyperphosphorylated during active transcription elongation (34,70) and that only hypophosphorylated Pol II can form the PIC to initiate transcription (30,66). These observations clearly indicate that CTD phosphorylation occurs between PIC formation and the transition from transcription initiation to elongation.…”

Section: Discussionmentioning

confidence: 55%

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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Section: Discussionmentioning

confidence: 55%

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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“…Previous work on the identity of the biological targets for acidic activators in vivo has suggested that these include TBP, TFIIB, and TAF40. Although TBP has been implicated as a target for several transcriptional activators (24,25,44), the fact the TBP also directly contacts a large number of TAFs and basal transcription factors, including RNA polymerase II and TFIIB, has led to some concern as to the in vivo relevance of these in vitro interactions (20,48,50). While TAF40 has been shown to contact the VP16 activation domain (15), this contact is to the C-terminal 452-to-490 sequence and not to the more N-terminal 412-to-456 sequence that, when multimerized, is fully sufficient to activate transcription efficiently (11,41).…”

Section: Discussionmentioning

confidence: 99%

Mutational analysis of the transcription activation domain of RelA: identification of a highly synergistic minimal acidic activation module.

Blair¹,

Bogerd²,

Madore³

et al. 1994

Mol. Cell. Biol.

107

124

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The potent C-terminal activation domain of the RelA(p65) subunit of the cellular transcription factor NF-KB is shown to contain several discrete acidic activation modules. These short, -11-amino-acid modules were able to give rise to only a low level of transcription activation when fused to the GAL4 DNA-binding domain as monomers. However, dimers and higher-order multimers activated the transcription of minimal promoter elements as effectively as the full-length RelA or VP16 activation domain. Therefore, this 11-aminoacid ReIA-derived acidic module appears to contain all of the sequence information required to fully activate a target promoter element as long as it is presented in a form that permits functional synergy. Critical primary sequence requirements for acidic activation module function included a core phenylalanine residue and flanking bulky hydrophobic residues. Overall negative charge was necessary but not sufficient for function. While dimeric forms of the 11-amino-acid acidic activation module bound to either TFIIB or TATA-binding protein efficiently in vitro, a similarly charged peptide lacking the core phenylalanine residue failed to interact.Overall, these data demonstrate that the biological activity of the RelA activation domain is dependent on acidic activator sequences that are closely comparable to those detected in the activation domain of the viral VP16 regulatory protein. We hypothesize that the ability of these acidic activators to specifically interact with multiple components of the transcription initiation complex likely underlies the dramatic functional synergy exhibited by this class of activation domains in vivo.

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“…25 bp upstream of the transcription initiation site. This element is an 8 bp consensus sequence (5h TATAAAA 3h), which is surrounded by GC-rich sequences and is recognized by TATA-binding protein (TBP) [33].…”

Section: Assembly Of the General Transcription Factors (Gtfs)mentioning

confidence: 99%

Transcriptional control and the role of silencers in transcriptional regulation in eukaryotes

Ogbourne

Antalis

1998

Biochemical Journal

219

171

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Mechanisms controlling transcription and its regulation are fundamental to our understanding of molecular biology and, ultimately, cellular biology. Our knowledge of transcription initiation and integral factors such as RNA polymerase is considerable, and more recently our understanding of the involvement of enhancers and complexes such as holoenzyme and mediator has increased dramatically. However, an understanding of transcriptional repression is also essential for a complete understanding of promoter structure and the regulation of gene expression. Transcriptional repression in eukaryotes is achieved through ‘silencers ’, of which there are two types, namely ‘silencer elements ’ and ‘negative regulatory elements ’ (NREs). Silencer elements are classical, position-independent elements that direct an active repression mechanism, and NREs are position-dependent elements that direct a passive repression mechanism. In addition, ‘repressors ’ are DNA-binding trasncription factors that interact directly with silencers. A review of the recent literature reveals that it is the silencer itself and its context within a given promoter, rather than the interacting repressor, that determines the mechanism of repression. Silencers form an intrinsic part of many eukaryotic promoters and, consequently, knowledge of their interactive role with enchancers and other transcriptional elements is essential for our understanding of gene regulation in eukaryotes.

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Specific interaction between the nonphosphorylated form of RNA polymerase II and the TATA-binding protein

Cited by 215 publications

References 42 publications

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

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

Mutational analysis of the transcription activation domain of RelA: identification of a highly synergistic minimal acidic activation module.

Transcriptional control and the role of silencers in transcriptional regulation in eukaryotes

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