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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...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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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. |
format | Online Article Text |
id | pubmed-6715403 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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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