The nsp1, nsp13, and M Proteins Contribute to the Hepatotropism of Murine Coronavirus JHM.WU
Identifieur interne : 000752 ( Pmc/Checkpoint ); précédent : 000751; suivant : 000753The nsp1, nsp13, and M Proteins Contribute to the Hepatotropism of Murine Coronavirus JHM.WU
Auteurs : Rong Zhang [États-Unis] ; Yize Li [États-Unis] ; Timothy J. Cowley [États-Unis] ; Adam D. Steinbrenner [États-Unis] ; Judith M. Phillips [États-Unis] ; Boyd L. Yount [États-Unis] ; Ralph S. Baric [États-Unis] ; Susan R. Weiss [États-Unis]Source :
- Journal of Virology [ 0022-538X ] ; 2015.
Abstract
Mouse hepatitis virus (MHV) isolates JHM.WU and JHM.SD promote severe central nervous system disease. However, while JHM.WU replicates robustly and induces hepatitis, JHM.SD fails to replicate or induce pathology in the liver. These two JHM variants encode homologous proteins with few polymorphisms, and little is known about which viral proteins(s) is responsible for the liver tropism of JHM.WU. We constructed reverse genetic systems for JHM.SD and JHM.WU and, utilizing these full-length cDNA clones, constructed chimeric viruses and mapped the virulence factors involved in liver tropism. Exchanging the spike proteins of the two viruses neither increased replication of JHM.SD in the liver nor attenuated JHM.WU. By further mapping, we found that polymorphisms in JHM.WU structural protein M and nonstructural replicase proteins nsp1 and nsp13 are essential for liver pathogenesis. M protein and nsp13, the helicase, of JHM.WU are required for efficient replication
Url:
DOI: 10.1128/JVI.03535-14
PubMed: 25589656
PubMed Central: 4403414
Affiliations:
- États-Unis
- Caroline du Nord, Pennsylvanie
- Chapel Hill (Caroline du Nord)
- Université de Caroline du Nord à Chapel Hill
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PMC:4403414Le document en format XML
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<front><div type="abstract" xml:lang="en"><title>ABSTRACT</title>
<p>Mouse hepatitis virus (MHV) isolates JHM.WU and JHM.SD promote severe central nervous system disease. However, while JHM.WU replicates robustly and induces hepatitis, JHM.SD fails to replicate or induce pathology in the liver. These two JHM variants encode homologous proteins with few polymorphisms, and little is known about which viral proteins(s) is responsible for the liver tropism of JHM.WU. We constructed reverse genetic systems for JHM.SD and JHM.WU and, utilizing these full-length cDNA clones, constructed chimeric viruses and mapped the virulence factors involved in liver tropism. Exchanging the spike proteins of the two viruses neither increased replication of JHM.SD in the liver nor attenuated JHM.WU. By further mapping, we found that polymorphisms in JHM.WU structural protein M and nonstructural replicase proteins nsp1 and nsp13 are essential for liver pathogenesis. M protein and nsp13, the helicase, of JHM.WU are required for efficient replication <italic>in vitro</italic>
and in the liver <italic>in vivo</italic>
. The JHM.SD nsp1 protein contains a K194R substitution of Lys194, a residue conserved among all other MHV strains. The K194R polymorphism has no effect on <italic>in vitro</italic>
replication but influences hepatotropism, and introduction of R194K into JHM.SD promotes replication in the liver. Conversely, a K194R substitution in nsp1 of JHM.WU or A59, another hepatotropic strain, significantly attenuates replication of each strain in the liver and increases IFN-β expression in macrophages in culture. Our data indicate that both structural and nonstructural proteins contribute to MHV liver pathogenesis and support previous reports that nsp1 is a <named-content content-type="genus-species">Betacoronavirus</named-content>
virulence factor.</p>
<p><bold>IMPORTANCE</bold>
The <named-content content-type="genus-species">Betacoronavirus</named-content>
genus includes human pathogens, some of which cause severe respiratory disease. The spread of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) into human populations demonstrates the zoonotic potential of emerging coronaviruses, and there are currently no vaccines or effective antivirals for human coronaviruses. Thus, it is important to understand the virus-host interaction that regulates coronavirus pathogenesis. Murine coronavirus infection of mice provides a useful model for the study of coronavirus-host interactions, including the determinants of tropism and virulence. We found that very small changes in coronavirus proteins can profoundly affect tropism and virulence. Furthermore, the hepatotropism of MHV-JHM depends not on the spike protein and viral entry but rather on a combination of the structural protein M and nonstructural replicase-associated proteins nsp1 and nsp13, which are conserved among betacoronaviruses. Understanding virulence determinants will aid in the design of vaccines and antiviral strategies.</p>
</div>
</front>
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<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Virol</journal-id>
<journal-id journal-id-type="hwp">jvi</journal-id>
<journal-id journal-id-type="pmc">jvi</journal-id>
<journal-id journal-id-type="publisher-id">JVI</journal-id>
<journal-title-group><journal-title>Journal of Virology</journal-title>
</journal-title-group>
<issn pub-type="ppub">0022-538X</issn>
<issn pub-type="epub">1098-5514</issn>
<publisher><publisher-name>American Society for Microbiology</publisher-name>
<publisher-loc>1752 N St., N.W., Washington, DC</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">25589656</article-id>
<article-id pub-id-type="pmc">4403414</article-id>
<article-id pub-id-type="publisher-id">03535-14</article-id>
<article-id pub-id-type="doi">10.1128/JVI.03535-14</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Pathogenesis and Immunity</subject>
</subj-group>
</article-categories>
<title-group><article-title>The nsp1, nsp13, and M Proteins Contribute to the Hepatotropism of Murine Coronavirus JHM.WU</article-title>
<alt-title alt-title-type="running-head">JHM Hepatotropism Maps to nsp1, nsp13, and M</alt-title>
<alt-title alt-title-type="short-authors">Zhang et al.</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Zhang</surname>
<given-names>Rong</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
<xref ref-type="author-notes" rid="fn1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Yize</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Cowley</surname>
<given-names>Timothy J.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
<xref ref-type="author-notes" rid="fn1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Steinbrenner</surname>
<given-names>Adam D.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
<xref ref-type="author-notes" rid="fn1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Phillips</surname>
<given-names>Judith M.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yount</surname>
<given-names>Boyd L.</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Baric</surname>
<given-names>Ralph S.</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>c</sup>
</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-8155-4528</contrib-id>
<name><surname>Weiss</surname>
<given-names>Susan R.</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
</contrib>
<aff id="aff1"><label>a</label>
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA</aff>
<aff id="aff2"><label>b</label>
Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA</aff>
<aff id="aff3"><label>c</label>
Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA</aff>
</contrib-group>
<contrib-group><contrib contrib-type="editor"><name><surname>Perlman</surname>
<given-names>S.</given-names>
</name>
<role>Editor</role>
</contrib>
</contrib-group>
<author-notes><corresp id="cor1">Address correspondence to Susan R. Weiss, <email>weisssr@upenn.edu</email>
.</corresp>
<fn id="fn1" fn-type="present-address"><label>*</label>
<p>Present address: Rong Zhang, Division of Infectious Diseases, School of Medicine, Washington University in St. Louis, St. Louis, Missouri, USA; Timothy J. Cowley, 13711 N.E. 10th Pl., Apt. 102, Bellevue, Washington, USA; Adam Steinbrenner, Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, California, USA.</p>
</fn>
<fn fn-type="equal"><p>R.Z. and Y.L. contributed equally to this article.</p>
</fn>
<fn fn-type="other"><p><bold>Citation</bold>
Zhang R, Li Y, Cowley TJ, Steinbrenner A, Phillips JM, Yount BL, Baric RS, Weiss SR. 2015. The nsp1, nsp13, and M proteins contribute to the hepatotropism of murine coronavirus JHM.WU. J Virol 89:3598–3609. doi:<ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1128/JVI.03535-14">10.1128/JVI.03535-14</ext-link>
.</p>
</fn>
</author-notes>
<pub-date pub-type="epub"><day>14</day>
<month>1</month>
<year>2015</year>
</pub-date>
<pub-date pub-type="collection"><day>1</day>
<month>4</month>
<year>2015</year>
</pub-date>
<volume>89</volume>
<issue>7</issue>
<fpage>3598</fpage>
<lpage>3609</lpage>
<history><date date-type="received"><day>8</day>
<month>12</month>
<year>2014</year>
</date>
<date date-type="accepted"><day>7</day>
<month>1</month>
<year>2015</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2015, American Society for Microbiology. All Rights Reserved.</copyright-statement>
<copyright-year>2015</copyright-year>
<copyright-holder>American Society for Microbiology</copyright-holder>
</permissions>
<self-uri content-type="pdf" xlink:href="zjv00715003598.pdf"></self-uri>
<abstract><title>ABSTRACT</title>
<p>Mouse hepatitis virus (MHV) isolates JHM.WU and JHM.SD promote severe central nervous system disease. However, while JHM.WU replicates robustly and induces hepatitis, JHM.SD fails to replicate or induce pathology in the liver. These two JHM variants encode homologous proteins with few polymorphisms, and little is known about which viral proteins(s) is responsible for the liver tropism of JHM.WU. We constructed reverse genetic systems for JHM.SD and JHM.WU and, utilizing these full-length cDNA clones, constructed chimeric viruses and mapped the virulence factors involved in liver tropism. Exchanging the spike proteins of the two viruses neither increased replication of JHM.SD in the liver nor attenuated JHM.WU. By further mapping, we found that polymorphisms in JHM.WU structural protein M and nonstructural replicase proteins nsp1 and nsp13 are essential for liver pathogenesis. M protein and nsp13, the helicase, of JHM.WU are required for efficient replication <italic>in vitro</italic>
and in the liver <italic>in vivo</italic>
. The JHM.SD nsp1 protein contains a K194R substitution of Lys194, a residue conserved among all other MHV strains. The K194R polymorphism has no effect on <italic>in vitro</italic>
replication but influences hepatotropism, and introduction of R194K into JHM.SD promotes replication in the liver. Conversely, a K194R substitution in nsp1 of JHM.WU or A59, another hepatotropic strain, significantly attenuates replication of each strain in the liver and increases IFN-β expression in macrophages in culture. Our data indicate that both structural and nonstructural proteins contribute to MHV liver pathogenesis and support previous reports that nsp1 is a <named-content content-type="genus-species">Betacoronavirus</named-content>
virulence factor.</p>
<p><bold>IMPORTANCE</bold>
The <named-content content-type="genus-species">Betacoronavirus</named-content>
genus includes human pathogens, some of which cause severe respiratory disease. The spread of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) into human populations demonstrates the zoonotic potential of emerging coronaviruses, and there are currently no vaccines or effective antivirals for human coronaviruses. Thus, it is important to understand the virus-host interaction that regulates coronavirus pathogenesis. Murine coronavirus infection of mice provides a useful model for the study of coronavirus-host interactions, including the determinants of tropism and virulence. We found that very small changes in coronavirus proteins can profoundly affect tropism and virulence. Furthermore, the hepatotropism of MHV-JHM depends not on the spike protein and viral entry but rather on a combination of the structural protein M and nonstructural replicase-associated proteins nsp1 and nsp13, which are conserved among betacoronaviruses. Understanding virulence determinants will aid in the design of vaccines and antiviral strategies.</p>
</abstract>
<counts><fig-count count="8"></fig-count>
<table-count count="1"></table-count>
<equation-count count="0"></equation-count>
<ref-count count="60"></ref-count>
<page-count count="12"></page-count>
<word-count count="9345"></word-count>
</counts>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>États-Unis</li>
</country>
<region><li>Caroline du Nord</li>
<li>Pennsylvanie</li>
</region>
<settlement><li>Chapel Hill (Caroline du Nord)</li>
</settlement>
<orgName><li>Université de Caroline du Nord à Chapel Hill</li>
</orgName>
</list>
<tree><country name="États-Unis"><region name="Pennsylvanie"><name sortKey="Zhang, Rong" sort="Zhang, Rong" uniqKey="Zhang R" first="Rong" last="Zhang">Rong Zhang</name>
</region>
<name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name>
<name sortKey="Baric, Ralph S" sort="Baric, Ralph S" uniqKey="Baric R" first="Ralph S." last="Baric">Ralph S. Baric</name>
<name sortKey="Cowley, Timothy J" sort="Cowley, Timothy J" uniqKey="Cowley T" first="Timothy J." last="Cowley">Timothy J. Cowley</name>
<name sortKey="Li, Yize" sort="Li, Yize" uniqKey="Li Y" first="Yize" last="Li">Yize Li</name>
<name sortKey="Phillips, Judith M" sort="Phillips, Judith M" uniqKey="Phillips J" first="Judith M." last="Phillips">Judith M. Phillips</name>
<name sortKey="Steinbrenner, Adam D" sort="Steinbrenner, Adam D" uniqKey="Steinbrenner A" first="Adam D." last="Steinbrenner">Adam D. Steinbrenner</name>
<name sortKey="Weiss, Susan R" sort="Weiss, Susan R" uniqKey="Weiss S" first="Susan R." last="Weiss">Susan R. Weiss</name>
<name sortKey="Yount, Boyd L" sort="Yount, Boyd L" uniqKey="Yount B" first="Boyd L." last="Yount">Boyd L. Yount</name>
</country>
</tree>
</affiliations>
</record>
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