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A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation

HIV-1 Tat stimulates transcription elongation by recruiting the P-TEFb (positive transcription elongation factor-b) (CycT1:CDK9) C-terminal domain (CTD) kinase to the HIV-1 promoter. Here we show that Tat transactivation also requires the Ssu72 CTD Ser5P (S5P)-specific phosphatase, which mediates tr...

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Autores principales: Chen, Yupeng, Zhang, Lirong, Estarás, Conchi, Choi, Seung H., Moreno, Luis, Karn, Jonathan, Moresco, James J., Yates, John R., Jones, Katherine A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4201287/
https://www.ncbi.nlm.nih.gov/pubmed/25319827
http://dx.doi.org/10.1101/gad.250449.114
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author Chen, Yupeng
Zhang, Lirong
Estarás, Conchi
Choi, Seung H.
Moreno, Luis
Karn, Jonathan
Moresco, James J.
Yates, John R.
Jones, Katherine A.
author_facet Chen, Yupeng
Zhang, Lirong
Estarás, Conchi
Choi, Seung H.
Moreno, Luis
Karn, Jonathan
Moresco, James J.
Yates, John R.
Jones, Katherine A.
author_sort Chen, Yupeng
collection PubMed
description HIV-1 Tat stimulates transcription elongation by recruiting the P-TEFb (positive transcription elongation factor-b) (CycT1:CDK9) C-terminal domain (CTD) kinase to the HIV-1 promoter. Here we show that Tat transactivation also requires the Ssu72 CTD Ser5P (S5P)-specific phosphatase, which mediates transcription termination and intragenic looping at eukaryotic genes. Importantly, HIV-1 Tat interacts directly with Ssu72 and strongly stimulates its CTD phosphatase activity. We found that Ssu72 is essential for Tat:P-TEFb-mediated phosphorylation of the S5P-CTD in vitro. Interestingly, Ssu72 also stimulates nascent HIV-1 transcription in a phosphatase-dependent manner in vivo. Chromatin immunoprecipitation (ChIP) experiments reveal that Ssu72, like P-TEFb and AFF4, is recruited by Tat to the integrated HIV-1 proviral promoter in TNF-α signaling 2D10 T cells and leaves the elongation complex prior to the termination site. ChIP-seq (ChIP combined with deep sequencing) and GRO-seq (genome-wide nuclear run-on [GRO] combined with deep sequencing) analysis further reveals that Ssu72 predominantly colocalizes with S5P–RNAPII (RNA polymerase II) at promoters in human embryonic stem cells, with a minor peak in the terminator region. A few genes, like NANOG, also have high Ssu72 at the terminator. Ssu72 is not required for transcription at most cellular genes but has a modest effect on cotranscriptional termination. We conclude that Tat alters the cellular function of Ssu72 to stimulate viral gene expression and facilitate the early S5P–S2P transition at the integrated HIV-1 promoter.
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spelling pubmed-42012872015-04-15 A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation Chen, Yupeng Zhang, Lirong Estarás, Conchi Choi, Seung H. Moreno, Luis Karn, Jonathan Moresco, James J. Yates, John R. Jones, Katherine A. Genes Dev Research Paper HIV-1 Tat stimulates transcription elongation by recruiting the P-TEFb (positive transcription elongation factor-b) (CycT1:CDK9) C-terminal domain (CTD) kinase to the HIV-1 promoter. Here we show that Tat transactivation also requires the Ssu72 CTD Ser5P (S5P)-specific phosphatase, which mediates transcription termination and intragenic looping at eukaryotic genes. Importantly, HIV-1 Tat interacts directly with Ssu72 and strongly stimulates its CTD phosphatase activity. We found that Ssu72 is essential for Tat:P-TEFb-mediated phosphorylation of the S5P-CTD in vitro. Interestingly, Ssu72 also stimulates nascent HIV-1 transcription in a phosphatase-dependent manner in vivo. Chromatin immunoprecipitation (ChIP) experiments reveal that Ssu72, like P-TEFb and AFF4, is recruited by Tat to the integrated HIV-1 proviral promoter in TNF-α signaling 2D10 T cells and leaves the elongation complex prior to the termination site. ChIP-seq (ChIP combined with deep sequencing) and GRO-seq (genome-wide nuclear run-on [GRO] combined with deep sequencing) analysis further reveals that Ssu72 predominantly colocalizes with S5P–RNAPII (RNA polymerase II) at promoters in human embryonic stem cells, with a minor peak in the terminator region. A few genes, like NANOG, also have high Ssu72 at the terminator. Ssu72 is not required for transcription at most cellular genes but has a modest effect on cotranscriptional termination. We conclude that Tat alters the cellular function of Ssu72 to stimulate viral gene expression and facilitate the early S5P–S2P transition at the integrated HIV-1 promoter. Cold Spring Harbor Laboratory Press 2014-10-15 /pmc/articles/PMC4201287/ /pubmed/25319827 http://dx.doi.org/10.1101/gad.250449.114 Text en © 2014 Chen et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Research Paper
Chen, Yupeng
Zhang, Lirong
Estarás, Conchi
Choi, Seung H.
Moreno, Luis
Karn, Jonathan
Moresco, James J.
Yates, John R.
Jones, Katherine A.
A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title_full A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title_fullStr A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title_full_unstemmed A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title_short A gene-specific role for the Ssu72 RNAPII CTD phosphatase in HIV-1 Tat transactivation
title_sort gene-specific role for the ssu72 rnapii ctd phosphatase in hiv-1 tat transactivation
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4201287/
https://www.ncbi.nlm.nih.gov/pubmed/25319827
http://dx.doi.org/10.1101/gad.250449.114
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