Linked Life Data

16154610

Source:http://linkedlifedata.com/resource/pubmed/id/16154610

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2005-11-15
pubmed:abstractText
Replication of the nonsegmented, plus-stranded RNA genome of Cucumber necrosis tombusvirus (CNV) requires two essential overlapping viral-coded replication proteins, the p33 replication co-factor and the p92 RNA-dependent RNA polymerase. In this paper, we demonstrate that p33 is phosphorylated in vivo and in vitro by a membrane-bound plant kinase. Phosphorylation of p33 was also demonstrated in vitro by using purified protein kinase C. The related p28 replication protein of Turnip crinkle virus was also found to be phosphorylated in vivo, suggesting that posttranslational modification of replication proteins is a general feature among members of the large Tombusviridae family. Based on in vitro studies with purified recombinant p33, we show evidence for phosphorylation of threonine and serine residues adjacent to the essential RNA-binding site in p33. Phosphorylation-mimicking aspartic acid mutations rendered p33 nonfunctional in plant protoplasts and in yeast, a model host. Comparable mutations within the prereadthrough portion of p92 did not abolish replication. The nonphosphorylation-mimicking alanine mutants of CNV were able to replicate in plant protoplasts and in yeast, albeit with reduced efficiency when compared to the wild type. These alanine mutants also showed altered subgenomic RNA synthesis and a reduction in the ratio between plus- and minus-strand RNAs produced during CNV infection. These findings suggest that phosphorylation of threonine/serine residues adjacent to the essential RNA-binding site in the auxiliary p33 protein likely plays a role in viral RNA replication and subgenomic RNA synthesis during tombusvirus infections.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0042-6822
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
343
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
65-78
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:16154610-Amino Acid Sequence, pubmed-meshheading:16154610-Amino Acid Substitution, pubmed-meshheading:16154610-Binding Sites, pubmed-meshheading:16154610-Blotting, Northern, pubmed-meshheading:16154610-Molecular Sequence Data, pubmed-meshheading:16154610-Mutation, Missense, pubmed-meshheading:16154610-Phosphorylation, pubmed-meshheading:16154610-Phosphotransferases, pubmed-meshheading:16154610-Plant Diseases, pubmed-meshheading:16154610-Protein Kinase C, pubmed-meshheading:16154610-Protein Processing, Post-Translational, pubmed-meshheading:16154610-RNA, Viral, pubmed-meshheading:16154610-RNA-Binding Proteins, pubmed-meshheading:16154610-RNA-Directed DNA Polymerase, pubmed-meshheading:16154610-Serine, pubmed-meshheading:16154610-Threonine, pubmed-meshheading:16154610-Tobacco, pubmed-meshheading:16154610-Tombusvirus, pubmed-meshheading:16154610-Viral Proteins, pubmed-meshheading:16154610-Virus Replication
pubmed:year
2005
pubmed:articleTitle
Phosphorylation of the p33 replication protein of Cucumber necrosis tombusvirus adjacent to the RNA binding site affects viral RNA replication.
pubmed:affiliation
Department of Plant Pathology, University of Kentucky, 201F Plant Science Building, Lexington, KY 40546, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural