SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b
Identifieur interne : 000734 ( Pmc/Checkpoint ); précédent : 000733; suivant : 000735SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b
Auteurs : Bhavna Varshney [Inde] ; Sudhakar Agnihotram [États-Unis] ; Yee-Joo Tan [Singapour] ; Ralph Baric [États-Unis] ; Sunil K. Lal [Inde]Source :
- PLoS ONE [ 1932-6203 ] ; 2012.
Abstract
The causative agent of severe acute respiratory syndrome, SARS coronavirus (SARS-CoV) genome encodes several unique group specific accessory proteins with unknown functions. Among them, accessory protein 3b (also known as ORF4) was lately identified as one of the viral interferon antagonist. Recently our lab uncovered a new role for 3b in upregulation of AP-1 transcriptional activity and its downstream genes. Thus, we believe that 3b might play an important role in SARS-CoV pathogenesis and therefore is of considerable interest. The current study aims at identifying novel host cellular interactors of the 3b protein.
In this study, using yeast two-hybrid and co-immunoprecipitation techniques, we have identified a host transcription factor RUNX1b (Runt related transcription factor, isoform b) as a novel interacting partner for SARS-CoV 3b protein. Chromatin immunoprecipitaion (ChIP) and reporter gene assays in 3b expressing jurkat cells showed recruitment of 3b on the RUNX1 binding element that led to an increase in RUNX1b transactivation potential on the IL2 promoter. Kinase assay and pharmacological inhibitor treatment implied that 3b also affect RUNX1b transcriptional activity by regulating its ERK dependent phosphorylation levels. Additionally, mRNA levels of MIP-1α, a RUNX1b target gene upregulated in SARS-CoV infected monocyte-derived dendritic cells, were found to be elevated in 3b expressing U937 monocyte cells.
These results unveil a novel interaction of SARS-CoV 3b with the host factor, RUNX1b, and speculate its physiological relevance in upregulating cytokines and chemokine levels in state of SARS virus infection.
Url:
DOI: 10.1371/journal.pone.0029542
PubMed: 22253733
PubMed Central: 3257236
Affiliations:
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>The causative agent of severe acute respiratory syndrome, SARS coronavirus (SARS-CoV) genome encodes several unique group specific accessory proteins with unknown functions. Among them, accessory protein 3b (also known as ORF4) was lately identified as one of the viral interferon antagonist. Recently our lab uncovered a new role for 3b in upregulation of AP-1 transcriptional activity and its downstream genes. Thus, we believe that 3b might play an important role in SARS-CoV pathogenesis and therefore is of considerable interest. The current study aims at identifying novel host cellular interactors of the 3b protein.</p>
</sec>
<sec><title>Methodology/Principal Findings</title>
<p>In this study, using yeast two-hybrid and co-immunoprecipitation techniques, we have identified a host transcription factor RUNX1b (Runt related transcription factor, isoform b) as a novel interacting partner for SARS-CoV 3b protein. Chromatin immunoprecipitaion (ChIP) and reporter gene assays in 3b expressing jurkat cells showed recruitment of 3b on the RUNX1 binding element that led to an increase in RUNX1b transactivation potential on the IL2 promoter. Kinase assay and pharmacological inhibitor treatment implied that 3b also affect RUNX1b transcriptional activity by regulating its ERK dependent phosphorylation levels. Additionally, mRNA levels of MIP-1α, a RUNX1b target gene upregulated in SARS-CoV infected monocyte-derived dendritic cells, were found to be elevated in 3b expressing U937 monocyte cells.</p>
</sec>
<sec><title>Conclusions/Significance</title>
<p>These results unveil a novel interaction of SARS-CoV 3b with the host factor, RUNX1b, and speculate its physiological relevance in upregulating cytokines and chemokine levels in state of SARS virus infection.</p>
</sec>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id>
<journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id>
<journal-id journal-id-type="publisher-id">plos</journal-id>
<journal-id journal-id-type="pmc">plosone</journal-id>
<journal-title-group><journal-title>PLoS ONE</journal-title>
</journal-title-group>
<issn pub-type="epub">1932-6203</issn>
<publisher><publisher-name>Public Library of Science</publisher-name>
<publisher-loc>San Francisco, USA</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">22253733</article-id>
<article-id pub-id-type="pmc">3257236</article-id>
<article-id pub-id-type="publisher-id">PONE-D-11-12481</article-id>
<article-id pub-id-type="doi">10.1371/journal.pone.0029542</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject>
</subj-group>
<subj-group subj-group-type="Discipline-v2"><subject>Biology</subject>
<subj-group><subject>Biochemistry</subject>
<subj-group><subject>Proteins</subject>
</subj-group>
</subj-group>
<subj-group><subject>Microbiology</subject>
<subj-group><subject>Virology</subject>
<subj-group><subject>Viral Classification</subject>
</subj-group>
</subj-group>
</subj-group>
<subj-group><subject>Molecular Cell Biology</subject>
<subj-group><subject>Signal Transduction</subject>
<subj-group><subject>Signaling Cascades</subject>
</subj-group>
</subj-group>
</subj-group>
<subj-group><subject>Proteomics</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group><article-title>SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b</article-title>
<alt-title alt-title-type="running-head">SARS-V 3b Protein Modulates RUNX1b Activity</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Varshney</surname>
<given-names>Bhavna</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Agnihotram</surname>
<given-names>Sudhakar</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Tan</surname>
<given-names>Yee-Joo</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Baric</surname>
<given-names>Ralph</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Lal</surname>
<given-names>Sunil K.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="corresp" rid="cor1"><sup>*</sup>
</xref>
</contrib>
</contrib-group>
<aff id="aff1"><label>1</label>
<addr-line>Virology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India</addr-line>
</aff>
<aff id="aff2"><label>2</label>
<addr-line>Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, United States of America</addr-line>
</aff>
<aff id="aff3"><label>3</label>
<addr-line>Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore</addr-line>
</aff>
<contrib-group><contrib contrib-type="editor"><name><surname>Sambhara</surname>
<given-names>Suryaprakash</given-names>
</name>
<role>Editor</role>
<xref ref-type="aff" rid="edit1"></xref>
</contrib>
</contrib-group>
<aff id="edit1">Centers for Disease Control and Prevention, United States of America</aff>
<author-notes><corresp id="cor1">* E-mail: <email>sunillal@icgeb.res.in</email>
</corresp>
<fn fn-type="con"><p>Conceived and designed the experiments: BV SA SL RB. Performed the experiments: BV SA RB SL. Analyzed the data: BV SL. Contributed reagents/materials/analysis tools: YT RB SL. Wrote the paper: BV SL.</p>
</fn>
</author-notes>
<pub-date pub-type="collection"><year>2012</year>
</pub-date>
<pub-date pub-type="epub"><day>12</day>
<month>1</month>
<year>2012</year>
</pub-date>
<volume>7</volume>
<issue>1</issue>
<elocation-id>e29542</elocation-id>
<history><date date-type="received"><day>1</day>
<month>7</month>
<year>2011</year>
</date>
<date date-type="accepted"><day>30</day>
<month>11</month>
<year>2011</year>
</date>
</history>
<permissions><copyright-statement>Varshney et al.</copyright-statement>
<copyright-year>2012</copyright-year>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.</license-p>
</license>
</permissions>
<abstract><sec><title>Background</title>
<p>The causative agent of severe acute respiratory syndrome, SARS coronavirus (SARS-CoV) genome encodes several unique group specific accessory proteins with unknown functions. Among them, accessory protein 3b (also known as ORF4) was lately identified as one of the viral interferon antagonist. Recently our lab uncovered a new role for 3b in upregulation of AP-1 transcriptional activity and its downstream genes. Thus, we believe that 3b might play an important role in SARS-CoV pathogenesis and therefore is of considerable interest. The current study aims at identifying novel host cellular interactors of the 3b protein.</p>
</sec>
<sec><title>Methodology/Principal Findings</title>
<p>In this study, using yeast two-hybrid and co-immunoprecipitation techniques, we have identified a host transcription factor RUNX1b (Runt related transcription factor, isoform b) as a novel interacting partner for SARS-CoV 3b protein. Chromatin immunoprecipitaion (ChIP) and reporter gene assays in 3b expressing jurkat cells showed recruitment of 3b on the RUNX1 binding element that led to an increase in RUNX1b transactivation potential on the IL2 promoter. Kinase assay and pharmacological inhibitor treatment implied that 3b also affect RUNX1b transcriptional activity by regulating its ERK dependent phosphorylation levels. Additionally, mRNA levels of MIP-1α, a RUNX1b target gene upregulated in SARS-CoV infected monocyte-derived dendritic cells, were found to be elevated in 3b expressing U937 monocyte cells.</p>
</sec>
<sec><title>Conclusions/Significance</title>
<p>These results unveil a novel interaction of SARS-CoV 3b with the host factor, RUNX1b, and speculate its physiological relevance in upregulating cytokines and chemokine levels in state of SARS virus infection.</p>
</sec>
</abstract>
<counts><page-count count="9"></page-count>
</counts>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>Inde</li>
<li>Singapour</li>
<li>États-Unis</li>
</country>
<region><li>Caroline du Nord</li>
</region>
<orgName><li>Université nationale de Singapour</li>
</orgName>
</list>
<tree><country name="Inde"><noRegion><name sortKey="Varshney, Bhavna" sort="Varshney, Bhavna" uniqKey="Varshney B" first="Bhavna" last="Varshney">Bhavna Varshney</name>
</noRegion>
<name sortKey="Lal, Sunil K" sort="Lal, Sunil K" uniqKey="Lal S" first="Sunil K." last="Lal">Sunil K. Lal</name>
</country>
<country name="États-Unis"><region name="Caroline du Nord"><name sortKey="Agnihotram, Sudhakar" sort="Agnihotram, Sudhakar" uniqKey="Agnihotram S" first="Sudhakar" last="Agnihotram">Sudhakar Agnihotram</name>
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<name sortKey="Baric, Ralph" sort="Baric, Ralph" uniqKey="Baric R" first="Ralph" last="Baric">Ralph Baric</name>
</country>
<country name="Singapour"><noRegion><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name>
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</tree>
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</record>
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