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Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein

Identifieur interne : 000176 ( Pmc/Curation ); précédent : 000175; suivant : 000177

Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein

Auteurs : Amruta S. Bhat ; H. S. Savithri

Source :

RBID : PMC:7086702

Abstract

Groundnut bud necrosis virus belongs to the genus Tospovirus, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in E. coli and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated in vitro by Nicotiana benthamiana plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family Bunyaviridae. The possible implications of the present findings for the viral life cycle are discussed.

Electronic supplementary material

The online version of this article (doi:10.1007/s00705-011-1110-0) contains supplementary material, which is available to authorized users.


Url:
DOI: 10.1007/s00705-011-1110-0
PubMed: 21947504
PubMed Central: 7086702

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PMC:7086702

Le document en format XML

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<p>Groundnut bud necrosis virus belongs to the genus
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<italic>E. coli</italic>
and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated
<italic>in vitro</italic>
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<italic>Nicotiana benthamiana</italic>
plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family
<italic>Bunyaviridae</italic>
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<pmc-dir>properties open_access</pmc-dir>
<front>
<journal-meta>
<journal-id journal-id-type="nlm-ta">Arch Virol</journal-id>
<journal-id journal-id-type="iso-abbrev">Arch. Virol</journal-id>
<journal-title-group>
<journal-title>Archives of Virology</journal-title>
</journal-title-group>
<issn pub-type="ppub">0304-8608</issn>
<issn pub-type="epub">1432-8798</issn>
<publisher>
<publisher-name>Springer Vienna</publisher-name>
<publisher-loc>Vienna</publisher-loc>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="pmid">21947504</article-id>
<article-id pub-id-type="pmc">7086702</article-id>
<article-id pub-id-type="publisher-id">1110</article-id>
<article-id pub-id-type="doi">10.1007/s00705-011-1110-0</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Original Article</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>Investigations on the RNA binding and phosphorylation of groundnut bud necrosis virus nucleocapsid protein</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Bhat</surname>
<given-names>Amruta S.</given-names>
</name>
<xref ref-type="aff" rid="Aff1"></xref>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Savithri</surname>
<given-names>H. S.</given-names>
</name>
<address>
<phone>+91-80-22932310</phone>
<fax>+91-80-23600814</fax>
<email>bchss@biochem.iisc.ernet.in</email>
</address>
<xref ref-type="aff" rid="Aff1"></xref>
</contrib>
<aff id="Aff1">
<institution-wrap>
<institution-id institution-id-type="GRID">grid.34980.36</institution-id>
<institution-id institution-id-type="ISNI">0000000104825067</institution-id>
<institution>Department of Biochemistry,</institution>
<institution>Indian Institute of Science,</institution>
</institution-wrap>
Bangalore, 560012 Karnataka State India</aff>
</contrib-group>
<pub-date pub-type="epub">
<day>24</day>
<month>9</month>
<year>2011</year>
</pub-date>
<pub-date pub-type="ppub">
<year>2011</year>
</pub-date>
<volume>156</volume>
<issue>12</issue>
<fpage>2163</fpage>
<lpage>2172</lpage>
<history>
<date date-type="received">
<day>17</day>
<month>4</month>
<year>2011</year>
</date>
<date date-type="accepted">
<day>6</day>
<month>9</month>
<year>2011</year>
</date>
</history>
<permissions>
<copyright-statement>© Springer-Verlag 2011</copyright-statement>
<license>
<license-p>This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.</license-p>
</license>
</permissions>
<abstract id="Abs1">
<p>Groundnut bud necrosis virus belongs to the genus
<italic>Tospovirus</italic>
, infects a wide range of crop plants and causes severe losses. To understand the role of the nucleocapsid protein in the viral life cycle, the protein was overexpressed in
<italic>E. coli</italic>
and purified by Ni-NTA chromatography. The purified N protein was well folded and was predominantly alpha-helical. Deletion analysis revealed that the C-terminal unfolded region of the N protein was involved in RNA binding. Furthermore, the N protein could be phosphorylated
<italic>in vitro</italic>
by
<italic>Nicotiana benthamiana</italic>
plant sap and by purified recombinant kinases such as protein kinase CK2 and calcium-dependent protein kinase. This is the first report of phoshphorylation of a nucleocapsid protein in the family
<italic>Bunyaviridae</italic>
. The possible implications of the present findings for the viral life cycle are discussed.</p>
<sec>
<title>Electronic supplementary material</title>
<p>The online version of this article (doi:10.1007/s00705-011-1110-0) contains supplementary material, which is available to authorized users.</p>
</sec>
</abstract>
<kwd-group xml:lang="en">
<title>Keywords</title>
<kwd>Infectious Bronchitis Virus</kwd>
<kwd>Tomato Spotted Wilt Virus</kwd>
<kwd>Nucleocapsid Protein</kwd>
<kwd>Viral Life Cycle</kwd>
<kwd>Rift Valley Fever</kwd>
</kwd-group>
<custom-meta-group>
<custom-meta>
<meta-name>issue-copyright-statement</meta-name>
<meta-value>© Springer-Verlag 2011</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
</front>
</pmc>
</record>

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