Transcription Elongation Activity of the Vaccinia Virus J3 Protein in Vivo Is Independent of Poly(A) Polymerase Stimulation
Identifieur interne : 002532 ( Istex/Curation ); précédent : 002531; suivant : 002533Transcription Elongation Activity of the Vaccinia Virus J3 Protein in Vivo Is Independent of Poly(A) Polymerase Stimulation
Auteurs : Ying Xiang ; Donald R. Latner ; Edward G. Niles ; Richard C. ConditSource :
- Virology [ 0042-6822 ] ; 2000.
English descriptors
- Teeft :
- Assay, Autoradiograms, Autoradiography, Biol, Brome mosaic virus, Chem, Codon, Condit, Confluent, Confluent monolayers, Deng, Edta, Elongation, Elongation factor activity, Equal volume, Final concentration, Gene, Gene product, Gershon, Host shutoff, Hybridization, Hybridized, Infection, Late rnas, Late times, Latner, Methyl groups, Methylation, Methyltransferase, Methyltransferase activity, Missense, Mops buffer, Mrna, Mutant, Mutant characterization, Mutant infections, Mutant rnas, Mutant transcripts, Mutant viruses, Mutation, Nacl, Ncoi, Normal amounts, Northern analysis, Nylon membranes, Oligonucleotide, Phenotype, Polyadenylation, Polymerase, Positive transcription elongation factor, Postinfection, Postreplicative, Promoter, Protein, Protein synthesis, Recombinant, Ribose, Ribose methylation, Rna, Rnase, Shuman, Size markers, Stimulatory, Stimulatory activities, Stimulatory activity, Subunit, Tail length, Time course, Transcript, Transcript release factor, Transcription, Transcription elongation, Vaccinia, Vaccinia gene, Vaccinia virus, Vaccinia virus gene, Various times, Various times postinfection, Viral, Virion, Virol, Virology, Virus, Western blot analysis, Xiang.
Abstract
Abstract: Prior genetic analysis suggests that the vaccinia virus J3 gene product, previously characterized as a bifunctional (nucleoside-2′-O-)-methyltransferase and poly(A) polymerase stimulatory factor, is a postreplicative positive transcription elongation factor. To test this hypothesis, viruses bearing mutations in the J3 gene were characterized with respect to viral protein and RNA synthesis in infected cells. The analysis reveals that compared to wt virus infections, J3 mutants synthesize reduced amounts of large late viral proteins and shorter-than-normal intermediate and late mRNAs. Structural analysis of one late mRNA shows that it is specifically truncated from the 3′ end, thus accounting for its shorter than normal chain length. Thus J3 mutant viruses are defective in elongation of transcription of postreplicative viral genes, strongly suggesting that the J3 gene product normally acts as a positive transcription elongation factor. Biochemical analysis of one J3 missense mutant demonstrates that it retains poly(A) stimulatory activity but is defective in (nucleoside-2′-O-)-methyltransferase activity. Thus the elongation factor activity of the J3 gene product is independent of the poly(A) stimulatory activity. It remains to be determined whether the (nucleoside-2′-O-)-methyltransferase and elongation factor activities of the J3 protein are linked or can be uncoupled by mutation.
Url:
DOI: 10.1006/viro.2000.0242
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Ying Xiang<affiliation><mods:affiliation>Department of Molecular Genetics and Microbiology, Center for Mammalian Genetics, University of Florida, Gainesville, Florida, 32610-0266</mods:affiliation>
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<affiliation><mods:affiliation>Department of Molecular Genetics and Microbiology, Center for Mammalian Genetics, University of Florida, Gainesville, Florida, 32610-0266</mods:affiliation>
<wicri:noCountry code="subField">32610-0266</wicri:noCountry>
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<affiliation><mods:affiliation>Department of Microbiology, Center for Microbial Pathogenesis, State University of New York, Buffalo, New York, 14214-3000</mods:affiliation>
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<term>Condit</term>
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<term>Confluent monolayers</term>
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<front><div type="abstract" xml:lang="en">Abstract: Prior genetic analysis suggests that the vaccinia virus J3 gene product, previously characterized as a bifunctional (nucleoside-2′-O-)-methyltransferase and poly(A) polymerase stimulatory factor, is a postreplicative positive transcription elongation factor. To test this hypothesis, viruses bearing mutations in the J3 gene were characterized with respect to viral protein and RNA synthesis in infected cells. The analysis reveals that compared to wt virus infections, J3 mutants synthesize reduced amounts of large late viral proteins and shorter-than-normal intermediate and late mRNAs. Structural analysis of one late mRNA shows that it is specifically truncated from the 3′ end, thus accounting for its shorter than normal chain length. Thus J3 mutant viruses are defective in elongation of transcription of postreplicative viral genes, strongly suggesting that the J3 gene product normally acts as a positive transcription elongation factor. Biochemical analysis of one J3 missense mutant demonstrates that it retains poly(A) stimulatory activity but is defective in (nucleoside-2′-O-)-methyltransferase activity. Thus the elongation factor activity of the J3 gene product is independent of the poly(A) stimulatory activity. It remains to be determined whether the (nucleoside-2′-O-)-methyltransferase and elongation factor activities of the J3 protein are linked or can be uncoupled by mutation.</div>
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