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Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb

The Positive Transcription Elongation Factor b (P-TEFb) phosphorylates Ser2 residues of the C-terminal domain (CTD) of the largest subunit (RPB1) of RNA polymerase II and is essential for the transition from transcription initiation to elongation in vivo. Surprisingly, P-TEFb exhibits Ser5 phosphory...

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Autores principales: Mayfield, Joshua E, Irani, Seema, Escobar, Edwin E, Zhang, Zhao, Burkholder, Nathaniel T, Robinson, Michelle R, Mehaffey, M Rachel, Sipe, Sarah N, Yang, Wanjie, Prescott, Nicholas A, Kathuria, Karan R, Liu, Zhijie, Brodbelt, Jennifer S, Zhang, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6715403/
https://www.ncbi.nlm.nih.gov/pubmed/31385803
http://dx.doi.org/10.7554/eLife.48725
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author Mayfield, Joshua E
Irani, Seema
Escobar, Edwin E
Zhang, Zhao
Burkholder, Nathaniel T
Robinson, Michelle R
Mehaffey, M Rachel
Sipe, Sarah N
Yang, Wanjie
Prescott, Nicholas A
Kathuria, Karan R
Liu, Zhijie
Brodbelt, Jennifer S
Zhang, Yan
author_facet Mayfield, Joshua E
Irani, Seema
Escobar, Edwin E
Zhang, Zhao
Burkholder, Nathaniel T
Robinson, Michelle R
Mehaffey, M Rachel
Sipe, Sarah N
Yang, Wanjie
Prescott, Nicholas A
Kathuria, Karan R
Liu, Zhijie
Brodbelt, Jennifer S
Zhang, Yan
author_sort Mayfield, Joshua E
collection PubMed
description The Positive Transcription Elongation Factor b (P-TEFb) phosphorylates Ser2 residues of the C-terminal domain (CTD) of the largest subunit (RPB1) of RNA polymerase II and is essential for the transition from transcription initiation to elongation in vivo. Surprisingly, P-TEFb exhibits Ser5 phosphorylation activity in vitro. The mechanism garnering Ser2 specificity to P-TEFb remains elusive and hinders understanding of the transition from transcription initiation to elongation. Through in vitro reconstruction of CTD phosphorylation, mass spectrometry analysis, and chromatin immunoprecipitation sequencing (ChIP-seq) analysis, we uncover a mechanism by which Tyr1 phosphorylation directs the kinase activity of P-TEFb and alters its specificity from Ser5 to Ser2. The loss of Tyr1 phosphorylation causes an accumulation of RNA polymerase II in the promoter region as detected by ChIP-seq. We demonstrate the ability of Tyr1 phosphorylation to generate a heterogeneous CTD modification landscape that expands the CTD’s coding potential. These findings provide direct experimental evidence for a combinatorial CTD phosphorylation code wherein previously installed modifications direct the identity and abundance of subsequent coding events by influencing the behavior of downstream enzymes.
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spelling pubmed-67154032019-09-03 Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb Mayfield, Joshua E Irani, Seema Escobar, Edwin E Zhang, Zhao Burkholder, Nathaniel T Robinson, Michelle R Mehaffey, M Rachel Sipe, Sarah N Yang, Wanjie Prescott, Nicholas A Kathuria, Karan R Liu, Zhijie Brodbelt, Jennifer S Zhang, Yan eLife Biochemistry and Chemical Biology The Positive Transcription Elongation Factor b (P-TEFb) phosphorylates Ser2 residues of the C-terminal domain (CTD) of the largest subunit (RPB1) of RNA polymerase II and is essential for the transition from transcription initiation to elongation in vivo. Surprisingly, P-TEFb exhibits Ser5 phosphorylation activity in vitro. The mechanism garnering Ser2 specificity to P-TEFb remains elusive and hinders understanding of the transition from transcription initiation to elongation. Through in vitro reconstruction of CTD phosphorylation, mass spectrometry analysis, and chromatin immunoprecipitation sequencing (ChIP-seq) analysis, we uncover a mechanism by which Tyr1 phosphorylation directs the kinase activity of P-TEFb and alters its specificity from Ser5 to Ser2. The loss of Tyr1 phosphorylation causes an accumulation of RNA polymerase II in the promoter region as detected by ChIP-seq. We demonstrate the ability of Tyr1 phosphorylation to generate a heterogeneous CTD modification landscape that expands the CTD’s coding potential. These findings provide direct experimental evidence for a combinatorial CTD phosphorylation code wherein previously installed modifications direct the identity and abundance of subsequent coding events by influencing the behavior of downstream enzymes. eLife Sciences Publications, Ltd 2019-08-06 /pmc/articles/PMC6715403/ /pubmed/31385803 http://dx.doi.org/10.7554/eLife.48725 Text en © 2019, Mayfield et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry and Chemical Biology
Mayfield, Joshua E
Irani, Seema
Escobar, Edwin E
Zhang, Zhao
Burkholder, Nathaniel T
Robinson, Michelle R
Mehaffey, M Rachel
Sipe, Sarah N
Yang, Wanjie
Prescott, Nicholas A
Kathuria, Karan R
Liu, Zhijie
Brodbelt, Jennifer S
Zhang, Yan
Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title_full Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title_fullStr Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title_full_unstemmed Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title_short Tyr1 phosphorylation promotes phosphorylation of Ser2 on the C-terminal domain of eukaryotic RNA polymerase II by P-TEFb
title_sort tyr1 phosphorylation promotes phosphorylation of ser2 on the c-terminal domain of eukaryotic rna polymerase ii by p-tefb
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6715403/
https://www.ncbi.nlm.nih.gov/pubmed/31385803
http://dx.doi.org/10.7554/eLife.48725
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