Heat-shock inactivation of the TFIIH-associated kinase and change in the phosphorylation sites on the C-terminal domain of RNA polymerase II

Dubois, Marie-Françoise; Vincent, Michel; Vigneron, Marc; Adamczewski, J P; Egly, Jean‐Marc; Bensaude, Olivier

doi:10.1093/nar/25.4.694

Cited by 53 publications

(52 citation statements)

References 76 publications

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“…However, preferential phosphorylation of heptapeptides with a lysine residue at position 7 over consensus heptapeptide sequences has been observed with synthetic CTD peptide substrates containing only four repeats (Rickert et al, 1999). In addition, while CDK7 CTD kinase activity is generally thought to be speci®c for Ser-5 (Sun et al, 1998;Trigon et al, 1998;Ramanathan et al, 2001), there are indications that Ser-2 phosphorylation by CDK7 might occur within less conserved heptapeptide repeats located at the C-terminal end of the CTD (Dubois et al, 1997;Bensaude et al, 1999). Results of kinetic studies with P-TEFb and puri®ed RNAP II as a substrate indicated slow initial phosphorylation events followed by more rapid phosphorylation of the CTD, suggesting that prior phosphorylation events can increase P-TEFb activity (Marshall et al, 1996).…”

Section: Distinct Ctd Kinase Speci®cities and Functionsmentioning

confidence: 99%

Regulation of RNA polymerase II activity by CTD phosphorylation and cell cycle control

Oelgeschläger

2002

Journal Cellular Physiology

127

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The carboxyl-terminal domain (CTD) of the largest subunit of mammalian RNA polymerase II (RNAP II) consists of 52 repeats of a consensus heptapeptide and is subject to phosphorylation and dephosphorylation events during each round of transcription. RNAP II activity is regulated during the cell cycle and cell cycledependend changes in RNAP II activity correlate well with CTD phosphorylation. In addition, global changes in the CTD phosphorylation status are observed in response to mitogenic or cytostatic signals such as growth factors, mitogens and DNA-damaging agents. Several CTD kinases are members of the cyclindependent kinase (CDK) superfamily and associate with transcription initiation complexes. Other CTD kinases implicated in cell cycle regulation include the mitogen-activated protein kinases ERK-1/2 and the c-Abl tyrosine kinase. These observations suggest that reversible RNAP II CTD phosphorylation may play a key role in linking cell cycle regulatory events to coordinated changes in transcription.

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Section: Distinct Ctd Kinase Speci®cities and Functionsmentioning

confidence: 99%

Regulation of RNA polymerase II activity by CTD phosphorylation and cell cycle control

Oelgeschläger

2002

Journal Cellular Physiology

127

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“…Treatment of cells with the transcription inhibitor actinomycin D or ␣-amanitin increases the ratio of RNAP IIO/IIA (46). Serum stimulation and heat shock increase the level of RNAP IIO via the cellular activation of MAPK2/ERK2 (47)(48)(49) and change the pattern of CTD phosphorylation (12)(13)(14). The increase in IIo signals, as indicated by increases in POL3/3 immunoreactivity relative to control extracts, confirms the above-mentioned studies that RNAP IIO levels are elevated in response to environmental stimuli (Fig.…”

Section: Fcp1 Dephosphorylates Phosphoserine 2 and Phosphoserine 5 Wimentioning

confidence: 99%

“…Serine 5 but not serine 2 phosphorylation recruits and activates the 5Ј-capping machinery (10, 11). Furthermore, nutritional stress and heat shock can independently alter the pattern of CTD phosphorylation, indicating that phosphoserine 2 and phosphoserine 5 are functionally different (12)(13)(14).A recent study using chromatin immunoprecipitation in Saccharomyces cerevisiae demonstrates that the pattern of CTD phosphorylation changes as RNAP II transcribes a given gene (15). Serine 5 phosphorylation is detected at the promoter regions, whereas serine 2 phosphorylation is increased as RNAP II leaves the promoter and transcribes the body of the gene.…”

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confidence: 99%

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TFIIF-associating Carboxyl-terminal Domain Phosphatase Dephosphorylates Phosphoserines 2 and 5 of RNA Polymerase II

Lin

Dubois

Dahmus

2002

Journal of Biological Chemistry

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The carboxyl-terminal domain (CTD) of the largest RNA polymerase (RNAP) II subunit undergoes reversible phosphorylation throughout the transcription cycle. The unphosphorylated form of RNAP II is referred to as IIA, whereas the hyperphosphorylated form is known as IIO. Phosphorylation occurs predominantly at serine 2 and serine 5 within the CTD heptapeptide repeat and has functional implications for RNAP II with respect to initiation, elongation, and transcription-coupled RNA processing. In an effort to determine the role of the major CTD phosphatase (FCP1) in regulating events in transcription that appear to be influenced by serine 2 and serine 5 phosphorylation, the specificity of FCP1 was examined. FCP1 is capable of dephosphorylating heterogeneous RNAP IIO populations of HeLa nuclear extracts. The extent of dephosphorylation at specific positions was assessed by immunoreactivity with monoclonal antibodies specific for phosphoserine 2 or phosphoserine 5. As an alternative method to assess FCP1 specificity, RNAP IIO isozymes were prepared in vitro by the phosphorylation of purified calf thymus RNAP IIA with specific CTD kinases and used as substrates for FCP1. FCP1 dephosphorylates serine 2 and serine 5 with comparable efficiency. Accordingly, the specificity of FCP1 is sufficiently broad to dephosphorylate RNAP IIO at any point in the transcription cycle irrespective of the site of serine phosphorylation within the consensus repeat. Reversible phosphorylation of the carboxyl-terminal domain (CTD)1 of the largest RNA polymerase (RNAP) II subunit plays an important role in the regulation of gene expression. The CTD of mammalian RNAP II is comprised of 52 repeats of the consensus sequence 1 YSPTSPS 7 (for a review, see Ref. 1). RNAP IIA, which contains an unmodified CTD, is actively recruited to the promoter as part of the preinitiation complex (2-5), whereas RNAP IIO, which contains a hyperphosphorylated CTD, is responsible for transcript elongation (6, 7). Therefore, protein kinases and phosphatases that alter the state of CTD phosphorylation can serve as transcriptional activators or repressors depending on the point in the transcription cycle at which they function.CTD phosphorylation occurs predominantly at serines 2 and 5 within the heptapeptide repeat. Genetic evidence indicates that the roles of serines in positions 2 and 5 are different. First, the partial substitution of serines in either position 2 or 5 have different effects on viability (8). Second, SRB (suppressors of RNA Polymerase IIB) mutations suppress the lethal effect of position 2 substitutions but not position 5 substitutions (9). Biochemical evidence has also confirmed differences between the two predominant serine positions. Serine 5 but not serine 2 phosphorylation recruits and activates the 5Ј-capping machinery (10, 11). Furthermore, nutritional stress and heat shock can independently alter the pattern of CTD phosphorylation, indicating that phosphoserine 2 and phosphoserine 5 are functionally different (12)(13)(14).A recent study using...

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“…Indeed, when TBP, which is essential for the assembly of the PIC, was omitted from the reaction mixture, CTD remained unphosphorylated (data not shown and Ref. 42). After classical in vitro transcription reaction performed in the presence of all the basal transcription factors as well as the four NTPs to allow the formation of the elongation complex, the phosphorylation of RNA pol II was visualized using an antibody raised against its large subunit (13).…”

Section: Fig 4 the Ring Finger Domain Of Mat1 Is Involved In Transcmentioning

confidence: 99%

Distinct Regions of MAT1 Regulate cdk7 Kinase and TFIIH Transcription Activities

Busso¹,

Keriel²,

Sandrock³

et al. 2000

Journal of Biological Chemistry

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The transcription/DNA repair factor TFIIH may be resolved into at least two subcomplexes: the core TFIIH and the cdk-activating kinase (CAK) complex. The CAK complex, which is also found free in the cell, is composed of cdk7, cyclin H, and MAT1. In the present work, we found that the C terminus of MAT1 binds to the cdk7⅐cyclin H complex and activates the cdk7 kinase activity. The median portion of MAT1, which contains a coiled-coil motif, allows the binding of CAK to the TFIIH core through interactions with both XPD and XPB helicases. Furthermore, using recombinant TFIIH complexes, it is demonstrated that the N-terminal RING finger domain of MAT1 is crucial for transcription activation and participates to the phosphorylation of the C-terminal domain of the largest subunit of the RNA polymerase II. Cyclin-dependent kinases (cdk)1 have a central role in the coordination of the eukaryotic cell cycle and participate in the integration of diverse growth regulatory signals. Some members of this protein kinase family are involved not only in cell cycle control but also in other mechanisms that govern cell life, notably transcription. Cdk activities are regulated by several different processes including the binding of an activating cyclin subunit, their phosphorylation, and the association of cdk inhibitors or other stimulatory factors (1). One of them, cdk7, was originally identified as the catalytic subunit of the cdk-activating kinase (CAK) complex that is able to phosphorylate, and thereby activate, several cdks (2). Besides cdk7, the CAK complex is composed of the regulatory subunit cyclin H (3, 4) and a third partner, MAT1, defined as a stabilizing factor (5, 6).Yeast genetic as well as biochemical studies demonstrated that MAT1 is also, together with cyclin H and cdk7, part of the general transcription factor TFIIH (7,8). TFIIH is composed of nine subunits and can be functionally divided into several subcomplexes such as the core TFIIH and the CAK complex (9, 10). TFIIH plays a role not only in transcription but also in DNA repair (11). Although TFIIH factor is absolutely required in nucleotide excision repair (11), the role of the CAK subcomplex in this reaction is not clear (12).Cdk7 phosphorylates several basal transcription factors (13-15) as well as the C-terminal domain (CTD) of the largest subunit of the RNA polymerase II (RNA pol II) (16). However, cdk7 is not the only kinase responsible for CTD phosphorylation. Indeed, the cdk8⅐cyclin C complex, which is found associated with the holoenzyme transcription complex, phosphorylates the CTD in vitro (17). Furthermore, the Srb10⅐Srb11 complex (the yeast counterpart of cdk8⅐cyclin C) phosphorylates the CTD in vivo prior to the formation of the preinitiation complex (PIC), resulting in an inhibition of the transcription reaction (18). Also, during human immunodeficiency virus infection, the CTD kinase activity of the cdk9⅐cyclin T complex, characterized as a component of the elongation factor pTEFb (for positive transcription elongation factor b), is required fo...

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Heat-shock inactivation of the TFIIH-associated kinase and change in the phosphorylation sites on the C-terminal domain of RNA polymerase II

Cited by 53 publications

References 76 publications

Regulation of RNA polymerase II activity by CTD phosphorylation and cell cycle control

Regulation of RNA polymerase II activity by CTD phosphorylation and cell cycle control

TFIIF-associating Carboxyl-terminal Domain Phosphatase Dephosphorylates Phosphoserines 2 and 5 of RNA Polymerase II

Distinct Regions of MAT1 Regulate cdk7 Kinase and TFIIH Transcription Activities

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