Linked Life Data

10873749

Source:http://linkedlifedata.com/resource/pubmed/id/10873749

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2000-7-19
pubmed:abstractText
Pyrazofurin (PZF), a cytidine analog and an inhibitor of orotate monophosphate decarboxylase, has been shown to decrease the levels of UTP and CTP in treated cells. When Sindbis virus (SV)-infected Aedes albopictus cells were treated with PZF, the yield of virus was reduced 100- to 1000-fold. By serial passage of our standard SV(STD) in Ae. albopictus cells in the presence of increasing concentrations of PZF, a mutant, SV(PZF), was derived, which was not inhibited by PZF. SV(PZF) is also resistant to adenosine, guanosine, and phosphono-acetyl-N-aspartate, all of which have been shown to decrease levels of UTP and CTP. Analysis of chimeric viruses containing sequences from the SV(PZF) and parental genomes showed that the sequence between nt 5262 and 7999 conferred the PZF-resistant phenotype. Sequencing of this region identified four mutations (nt 5750, 6627, 7543, and 7593), which are predicted to lead to amino acid changes: opal550L in nsP3 and M287L, K592I, and P609T in nsP4. Characterization of viruses containing one or more of these mutations demonstrated that all three mutations in the nsP4 coding region are required to produce full resistance to PZF. Using a molecular model of nsP4 based on the structure of HIV reverse transcriptase, we located amino acid change M287L at the tip of the fingers domain and K592I and P609T at the base of the thumb domain of the viral RNA polymerase. We suggest that these three amino acid changes in nsP4 alter the geometry of the NTP binding pocket so as to increase the affinity of the enzyme for CTP and UTP.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0042-6822
pubmed:author
pubmed:copyrightInfo
Copyright 2000 Academic Press.
pubmed:issnType
Print
pubmed:day
20
pubmed:volume
272
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
61-71
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:10873749-Adenosine, pubmed-meshheading:10873749-Aedes, pubmed-meshheading:10873749-Amino Acid Substitution, pubmed-meshheading:10873749-Animals, pubmed-meshheading:10873749-Aspartic Acid, pubmed-meshheading:10873749-Binding Sites, pubmed-meshheading:10873749-Cells, Cultured, pubmed-meshheading:10873749-Chick Embryo, pubmed-meshheading:10873749-DNA-Directed RNA Polymerases, pubmed-meshheading:10873749-Dose-Response Relationship, Drug, pubmed-meshheading:10873749-Drug Resistance, Microbial, pubmed-meshheading:10873749-Fibroblasts, pubmed-meshheading:10873749-Guanosine, pubmed-meshheading:10873749-Models, Molecular, pubmed-meshheading:10873749-Mutation, pubmed-meshheading:10873749-Phenotype, pubmed-meshheading:10873749-Phosphonoacetic Acid, pubmed-meshheading:10873749-Protein Structure, Tertiary, pubmed-meshheading:10873749-Recombination, Genetic, pubmed-meshheading:10873749-Ribonucleosides, pubmed-meshheading:10873749-Sindbis Virus, pubmed-meshheading:10873749-Virus Replication
pubmed:year
2000
pubmed:articleTitle
A mutant of Sindbis virus that is resistant to pyrazofurin encodes an altered RNA polymerase.
pubmed:affiliation
Department of Molecular Genetics and Microbiology, UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey, 08854-5635, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't