Abstract:
:The phosphoprotein (P protein) of respiratory syncytial virus (RSV) is a key component of the viral RNA-dependent RNA polymerase complex. The protein is constitutively phosphorylated at the two clusters of serine residues (116, 117, and 119 [116/117/119] and 232 and 237 [232/237]). To examine the role of phosphorylation of the RSV P protein in virus replication, these five serine residues were altered to eliminate their phosphorylation potential, and the mutant proteins were analyzed for their functions with a minigenome assay. The reporter gene expression was reduced by 20% when all five phosphorylation sites were eliminated. Mutants with knockout mutations at two phosphorylation sites (S232A/S237A [PP2]) and at five phosphorylation sites (S116L/S117R/S119L/S232A/S237A [PP5]) were introduced into the infectious RSV A2 strain. Immunoprecipitation of (33)P(i)-labeled infected cells showed that P protein phosphorylation was reduced by 80% for rA2-PP2 and 95% for rA2-PP5. The interaction between the nucleocapsid (N) protein and P protein was reduced in rA2-PP2- and rA2-PP5-infected cells by 30 and 60%, respectively. Although the two recombinant viruses replicated well in Vero cells, rA2-PP2 and, to a greater extent, rA2-PP5, replicated poorly in HEp-2 cells. Virus budding from the infected HEp-2 cells was affected by dephosphorylation of P protein, because the majority of rA2-PP5 remained cell associated. In addition, rA2-PP5 was also more attenuated than rA2-PP2 in replication in the respiratory tracts of mice and cotton rats. Thus, our data suggest that although the major phosphorylation sites of RSV P protein are dispensable for virus replication in vitro, phosphorylation of P protein is required for efficient virus replication in vitro and in vivo.
journal_name
J Viroljournal_title
Journal of virologyauthors
Lu B,Ma CH,Brazas R,Jin Hdoi
10.1128/jvi.76.21.10776-10784.2002subject
Has Abstractpub_date
2002-11-01 00:00:00pages
10776-84issue
21eissn
0022-538Xissn
1098-5514journal_volume
76pub_type
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abstract::A DNA vaccine plasmid containing the Japanese encephalitis (JE) virus premembrane (prM) and envelope (E) genes (designated pcDNA3JEME) was evaluated for immunogenicity and protective efficacy in mice. Two immunizations of 4-week-old female ICR mice with pcDNA3JEME by intramuscular or intradermal injections at a dose o...
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pub_type: 杂志文章
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pub_type: 临床试验,杂志文章
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更新日期:1999-01-01 00:00:00
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pub_type: 杂志文章
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更新日期:1985-07-01 00:00:00
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doi:10.1128/JVI.68.12.7709-7716.1994
更新日期:1994-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:1982-12-01 00:00:00
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journal_title:Journal of virology
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更新日期:1991-08-01 00:00:00
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pub_type: 杂志文章,评审
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pub_type: 杂志文章
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更新日期:2007-10-01 00:00:00
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更新日期:2005-09-01 00:00:00
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pub_type: 杂志文章
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更新日期:1991-04-01 00:00:00
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更新日期:2001-02-01 00:00:00