Breaking barriers to transcription elongation

Saunders, Abbie; Core, Leighton J.; Lis, John T.

doi:10.1038/nrm1981

Cited by 447 publications

(532 citation statements)

References 151 publications

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“…GST-(YSPTSPS) [8] GST M kDa 97 66 45 30 20 GST-p(YSPTSPS) [8] Coomassie A GsT-CTD [13] fusion protein containing 13 hepta-repeats (10 µm) was incubated with 0.1 µm P-TEFb and 300 µm [ 32 P]-γ-labelled ATP. saturation of the phosphorylation reaction was achieved within 3 h. Data represent the mean ± range from two independent experiments.…”

Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

“…(b) sDs-PAGE analysis of CTD phosphorylation by P-TEFb. At concentrations of 0.1 µm P-TEFb, 100 µm GsT-CTD [8] and 3 mm ATP the substrate was fully phosphorylated after 4 h at 30 °C. (c) EsI-ms analysis of CTD phosphorylation in a time course experiment.…”

Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

“…shown are time points at the beginning and after 1, 2 and 16 h of the reaction. For the last time point, a GsT-CTD [8] substrate was used for better ionization properties in the EsI-ms analysis.…”

Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

“…Using the in vitro kinase assay, we set out to determine the molecular requirements of P-TEFb inhibition quantitatively. Under standard conditions, 0.1 µM P-TEFb was mixed with 100 µM GST-CTD [8] and 100 µM ATP and incubated for 10 min before the solution was spotted onto filter paper and the reaction stopped by immersing the filter paper in phosphoric acid. Under these conditions full-length Hexim1 at a concentration of 2.5 µM inhibited P-TEFb activity to 28% without addition of 7SK snRNA.…”

Section: S S S Y S P T P S Y S P T P S Y S P T P Smentioning

confidence: 99%

“…In humans, the CTD comprises 52 repeats of the consensus sequence Y 1 S 2 P 3 T 4 S 5 P 6 S 7 that exhibits only some variations from the strict consensus towards the C-terminus [3][4][5][6][7] . Phosphorylation of Ser5 by Cdk7 and Cdk8 kinases of TFIIH and the mediator complex, respectively, has been described to be concomitant with transcription initiation, whereas Cdk9 of P-TEFb is suggested to phosphorylate Ser2, which marks the elongation phase of transcription [3][4][5][6][7][8] . In addition, Ser7 of the CTD can also be phosphorylated, which has been linked to small nuclear RNAs (snRNA) transcription and recruitment of the integrator complex [9][10][11][12][13] .…”

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Serine-7 but not serine-5 phosphorylation primes RNA polymerase II CTD for P-TEFb recognition

2012

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Phosphorylation of RnA polymerase II carboxy-terminal domain (CTD) in hepta-repeats YsPTsPs regulates eukaryotic transcription. Whereas ser5 is phosphorylated in the initiation phase, ser2 phosphorylation marks the elongation state. Here we show that the positive transcription elongation factor P-TEFb is a ser5 CTD kinase that is unable to create ser2/ser5 double phosphorylations, while it exhibits fourfold higher activity on a CTD substrate prephosphorylated at ser7 compared with the consensus hepta-repeat or the YsPTsPK variant. mass spectrometry reveals an equal number of phosphorylations to the number of heptarepeats provided, yet the mechanism of phosphorylation is distributive despite the repetitive nature of the substrate. Inhibition of P-TEFb activity is mediated by two regions in Hexim1 that act synergistically on Cdk9 and Cyclin T1. HIV-1 Tat/TAR abrogates Hexim1 inhibition to stimulate transcription of viral genes but does not change the substrate specificity. Together, these results provide insight into the multifaceted pattern of CTD phosphorylation.

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Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

Section: Reverse Kinase Order Does Not Induce Double Phosphorylationsmentioning

confidence: 99%

Section: S S S Y S P T P S Y S P T P S Y S P T P Smentioning

confidence: 99%

mentioning

confidence: 99%

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Serine-7 but not serine-5 phosphorylation primes RNA polymerase II CTD for P-TEFb recognition

2012

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Epigenetic Histone Changes in the Toxicologic Mode of Action of Arsenic

Reichard

Puga

2012

Toxicology and Epigenetics

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Transcriptional silencing

2015

The Dictionary of Genomics, Transcriptomics and Proteomics

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Silent chromatin in budding yeast is characterized by the presence of a specialized chromatin modification complex consisting of silent information regulator (Sir) proteins, closely packed pairs of nucleosomes, and hypoacetylated and hypomethylated histones. How this specialized chromatin is established, maintained and inherited has been extensively studied. Less investigated are the determinants that constrain its linear spread along the chromatin fibre and the manner by which it represses gene transcription. Here we review the essential features of SIR-mediated heterochromatin, and discuss genomic and proteomic approaches for discerning the composition of its boundaries and for elucidating the mechanisms by which it silences transcription. Lu Gao is an orthopaedic surgeon. He is also a predoctoral student of Biochemistry and Molecular Biology, and is studying mechanisms of epigenetic gene silencing for his PhD degree. David S. Gross is a professor at the Louisiana State University Health Sciences Center. He is interested in mechanisms by which chromatin regulates gene expression. Current projects in his lab are focused on transcriptional silencing, heat shock gene regulation and p53 transcriptional regulatory mechanisms.

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Breaking barriers to transcription elongation

Cited by 447 publications

References 151 publications

Serine-7 but not serine-5 phosphorylation primes RNA polymerase II CTD for P-TEFb recognition

Serine-7 but not serine-5 phosphorylation primes RNA polymerase II CTD for P-TEFb recognition

Epigenetic Histone Changes in the Toxicologic Mode of Action of Arsenic

Transcriptional silencing

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