A protein phosphatase functions to recycle RNA polymerase II

Cho, Helen; Kim, Tae Kyung; Mancebo, Helena; Lane, William S.; Flores, Osvaldo; Reinberg, Danny

doi:10.1101/gad.13.12.1540

Cited by 179 publications

(239 citation statements)

References 46 publications

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“…Serine͞ threonine protein phosphatases have been divided into four families on the basis of biochemical properties and amino acid sequence similarity (41). Both the HeLa cell and yeast CTD phosphatases have been classified as type 2C phosphatases on the strength of their requirement for magnesium ions and resistance to okadaic acid (24,42). It is remarkable, therefore, that our hRAP74cc structure similarity search identified a discrete HTH domain [Dali server (37) Z score ϭ 3.1, ␣-carbon superposition rmsd ϭ 2.2 Å] located at the C terminus of another phosphatase 2C family member, protein phosphatase 2C (PP2C) (43), which catalyzes dephosphorylation of AMP-activated protein kinase C (44).…”

Section: Resultsmentioning

confidence: 99%

“…The activity of this HeLa cell CTD phosphatase is stimulated by the RAP74 subunit of TFIIF, and this stimulation can be inhibited by TFIIB (22). A yeast two-hybrid screen for RAP74-interacting proteins produced partial cDNAs encoding a human CTD phosphatase, FCP1 (23,24). The C-terminal region (residues 436-517) of human RAP74, which interacts with the FCP1 phosphatase, is both necessary and sufficient for RAP74-mediated stimulation of CTD phosphatase activity in vitro (23).…”

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confidence: 99%

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Crystal structure of the C-terminal domain of the RAP74 subunit of human transcription factor IIF

Kamada

Angelis

Roeder

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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The x-ray structure of a C-terminal fragment of the RAP74 subunit of human transcription factor (TF) IIF has been determined at 1.02-Å resolution. The ␣͞␤ structure is strikingly similar to the globular domain of linker histone H5 and the DNA-binding domain of hepatocyte nuclear factor 3␥ (HNF-3␥), making it a winged-helix protein. The surface electrostatic properties of this compact domain differ significantly from those of bona fide winged-helix transcription factors (HNF-3␥ and RFX1) and from the winged-helix domains found within the RAP30 subunit of TFIIF and the ␤ subunit of TFIIE. RAP74 has been shown to interact with the TFIIF-associated C-terminal domain phosphatase FCP1, and a putative phosphatase binding site has been identified within the RAP74 wingedhelix domain.T ranscription of class II nuclear genes in eukaryotes requires a complex assembly of proteins composed of a multisubunit RNA polymerase II (pol II) and general transcription factors (TFIIs), which are required for assembly of the preinitiation complex (PIC) on promoter DNA and initiation of transcription (1). In the most general case, PIC assembly begins with the TATA box-binding protein (TBP) subunit of TFIID recognizing the TATA element, followed by coordinated accretion of TFIIB, the nonphosphorylated form of pol II plus TFIIF, TFIIE, and TFIIH. Transcription initiation is followed by phosphorylation of the C-terminal domain (CTD) of the largest subunit of pol II, which in turn facilitates promoter clearance and productive elongation. After elongation and termination of pre-mRNA synthesis, a phosphatase recycles pol II to its nonphosphorylated form, allowing the enzyme and the requisite TFIIs to initiate the next round of transcription.Mammalian TFIIF is an ␣␤ hetero-oligomer of RAP74 and RAP30 subunits (RNA polymerase II-associating proteins; 58 kDa and 28 kDa, respectively) (2), which binds to pol II in solution and facilitates delivery of the polymerase to the TFIID-TFIIB-DNA complex on the promoter. TFIIF was originally identified on the basis of physical association with pol II, and its requirement for promoter-specific transcription initiation by this large multisubunit enzyme. Within the PIC, TFIIF stabilizes binding of pol II to TFIID and TFIIB at the promoter (3-5). Moreover, TFIIF is required for entry of TFIIE and TFIIH into the PIC, for subsequent open complex formation catalyzed by the DNA helicase subunit of TFIIH (5-7), and for synthesis of the first phosphodiester bond of nascent transcripts (8-10). It is also possible that TFIIF acts as a scaffold for binding of TFIIH to the growing PIC and͞or that TFIIF actively participates in formation of the open complex. Photocrosslinking experiments suggest that TFIIF interacts with promoter DNA to induce a dramatic conformational change that results in wrapping of DNA around pol II and the class II general transcription factors within the PIC (11-14).Biochemical properties of TFIIF have been attributed to distinct polypeptide chain segments of each of subunit. Mammalian RAP30 can b...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Crystal structure of the C-terminal domain of the RAP74 subunit of human transcription factor IIF

Kamada

Angelis

Roeder

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, Fcp1 dephosphorylates Ser2 preferentially while Ssu72 (SCP1 in humans), which is a component of the yeast 39-end machinery, dephosphorylates Ser5 (Meinhart et al 2005). Dephosphorylation of Ser2 is also critical for RNAPII recycling (Cho et al 1999). Both unphosphorylated and Ser2 phosphorylated CTD accumulate near transcription termination sites, suggesting that CTD dephosphorylation occurs during or prior to RNAPII release (Lian et al 2008).…”

Section: Ctd Of the Rnapii Large Subunitmentioning

confidence: 99%

Transcription termination by nuclear RNA polymerases

Richard

Manley²

2009

Genes Dev.

291

326

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Gene transcription in the cell nucleus is a complex and highly regulated process. Transcription in eukaryotes requires three distinct RNA polymerases, each of which employs its own mechanisms for initiation, elongation, and termination. Termination mechanisms vary considerably, ranging from relatively simple to exceptionally complex. In this review, we describe the present state of knowledge on how each of the three RNA polymerases terminates and how mechanisms are conserved, or vary, from yeast to human.

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“…It has been purified from yeast and mammalian cells (8,9), and the corresponding cDNA has been cloned by a two-hybrid screen using the RAP74 subunit of the general transcription factor TFIIF as a bait (10). FCP1 (TFIIF-dependent CTD Phosphatase 1) consists of a single 150-kDa polypeptide subunit (11,12). FCP1…”

Section: Rna Polymerase II (Rnap Ii)mentioning

confidence: 99%

FCP1 Phosphorylation by Casein Kinase 2 Enhances Binding to TFIIF and RNA Polymerase II Carboxyl-terminal Domain Phosphatase Activity

Palancade

Dubois

Bensaude

2002

Journal of Biological Chemistry

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Dephosphorylation of RNA polymerase II carboxylterminal domain (CTD) is required to resume sequential transcription cycles. FCP1 (TFIIF-dependent CTD phosphatase 1) is the only known phosphatase targeting RNAP II CTD. Here we show that in Xenopus laevis cells, xFCP1 is a phosphoprotein. On the basis of biochemical fractionation and drug sensitivity, casein kinase 2 (CK2) is shown to be the major kinase involved in xFCP1 phosphorylation in X. laevis egg extracts. CK2 phosphorylates xFCP1 mainly at a cluster of serines centered on Ser 457 . CK2-dependent phosphorylation enhances 4-fold the CTD phosphatase activity of FCP1 and its binding to the RAP74 subunit of general transcription factor TFIIF. These findings unravel a new mechanism regulating CTD phosphorylation and hence class II gene transcription.RNA polymerase II (RNAP II) 1 is a multiprotein complex in charge of eucaryotic mRNA synthesis (1). Rpb1, the largest RNAP II subunit, is extensively phosphorylatable on its carboxyl-terminal domain (CTD), which consists of up to 52 repeats of the consensus heptapeptide Tyr-Ser-Pro-Thr-Ser-ProSer (2). Two phosphoisoforms of RNAP II coexist in somatic cells in a dynamic equilibrium (3). The hypophosphorylated IIA form assembles into preinitiation complexes, whereas the hyperphosphorylated IIO form elongates mRNA synthesis and recruits mRNA processing factors (4). The conversion from IIA to IIO occurs during initiation and elongation of transcription and involves CTD kinases such as CDK7 and CDK9, respectively, included in the transcription factors TFIIH and p-TEFb (5, 6). CTD dephosphorylation is required to recycle RNAP II after a transcriptional round (7).In contrast to the numerous CTD kinases identified, only one CTD phosphatase has been characterized so far. It has been purified from yeast and mammalian cells (8,9), and the corresponding cDNA has been cloned by a two-hybrid screen using the RAP74 subunit of the general transcription factor TFIIF as a bait (10). FCP1 (TFIIF-dependent CTD Phosphatase 1) consists of a single 150-kDa polypeptide subunit (11, 12). FCP1does not display any homology with other known protein phosphatases but possesses an unusual catalytic site present in a subset of phosphotransferases (13,14). FCP1 is a highly specific phosphatase as the CTD within a functional RNAP II remains its only known protein substrate. FCP1 is recruited to RNAP II through docking sites on the Rpb4 subunit of RNAP II and on the RAP74 subunit of TFIIF (15, 16).FCP1 activity is regulated at various levels. First, competitive binding of general transcription factors TFIIF and TFIIB to FCP1 carboxyl-terminal domain, respectively, stimulates or inhibits its phosphatase activity (15,17). Second, free RNAP IIO molecules are better substrates than molecules engaged in transcription elongation complexes (18,19). Third, inhibition of FCP1 phosphatase activity by the human immunodeficiency viral protein Tat may contribute to stimulate transcriptional elongation from the human immunodeficiency virus promoter (20, 21).We had...

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A protein phosphatase functions to recycle RNA polymerase II

Cited by 179 publications

References 46 publications

Crystal structure of the C-terminal domain of the RAP74 subunit of human transcription factor IIF

Crystal structure of the C-terminal domain of the RAP74 subunit of human transcription factor IIF

Transcription termination by nuclear RNA polymerases

FCP1 Phosphorylation by Casein Kinase 2 Enhances Binding to TFIIF and RNA Polymerase II Carboxyl-terminal Domain Phosphatase Activity

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