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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2014
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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. |
format | Online Article Text |
id | pubmed-4201287 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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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