Phosphorylation Generates Different Forms of Rotavirus NSP5

Afrikanova, Ivka; Miozzo, Maria C.; Giambiagi, Susana; Burrone, Óscar R.

doi:10.1099/0022-1317-77-9-2059

Cited by 72 publications

(107 citation statements)

References 22 publications

(21 reference statements)

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“…The last 18 C-terminal aa are highly conserved even in group C viruses (Mattion et al, 1991). We have recently shown that NSP5 is hyperphosphorylated in vivo through a complex process, involving autophosphorylation, which gives I. Afrikanova and others I. Afrikanova and others rise to a number of different isoforms (26, 28 and 32-34 kDa) (Afrikanova et al, 1996). Similar results have also been obtained by other authors (Blackhall et al, 1997 ;Poncet et al, 1997).…”

Section: Introductionsupporting

confidence: 79%

“…We used DSP as a chemical cross-linker since it permeates into living cells, is able to cross-link proteins interacting at distances of up to 12 A H , and is sensitive to reducing agents. Analysis of the immunoprecipitates derived from cross-linked virusinfected cell extracts showed two new proteins in addition to CGIA Rotavirus NSP2 up-regulates NSP5 phosphorylation Rotavirus NSP2 up-regulates NSP5 phosphorylation the different phosphorylated isoforms of NSP5 (Afrikanova et al, 1996). These two proteins migrated on SDS-PAGE identically to the viral proteins VP1 and NSP2 of molecular masses 120 and 35 kDa, respectively ( Fig.…”

Section: In Vivo Chemical Cross-linkingmentioning

confidence: 97%

“…Immunoprecipitation experiments were performed as described by Afrikanova et al (1996). For immunoblots, lysates of transfected or cotransfected cells (corresponding to 0n5i10& cells) were resolved on 12 % SDS-polyacrylamide gels (Laemmli et al, 1970), transferred onto a PVDF (Millipore) membrane and reacted to a guinea pig anti-NSP5 serum.…”

Section: Immunoprecipitation and Western Immunoblot Analysismentioning

confidence: 99%

“…In vitro phosphorylation was carried out essentially as described by Afrikanova et al (1996) by incubating Protein A-Sepharose beads (Pharmacia) containing immunoprecipitates with [γ-$#P]ATP (Amersham) for 30 min at 37 mC in kinase buffer (50 mM Tris-HCl, pH 8n0, 1n5 mM spermidine, 5 mM MgCl # , 1 mM DTT, 5 % glycerol). Phosphatase treatments were performed as described (Afrikanova et al, 1996). The immunoprecipitates were eluted by 4i Laemmli's sample buffer at 96 mC for 3 min.…”

Section: Radiolabelling With [ 35 S]methionine Immunoprecipitationmentioning

confidence: 99%

“…Labelling of cells with [$&S]methionine was performed essentially as described by Afrikanova et al (1996). Total cellular extracts were prepared in TNN buffer (100 mM Tris-HCl, pH 8n0, 250 mM NaCl, 0n5 % NP40, 1 mM PMSF) and used for immunoprecipitation and PAGE analysis as described (Afrikanova et al, 1996).…”

Section: Radiolabelling With [ 35 S]methionine Immunoprecipitationmentioning

confidence: 99%

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Rotavirus NSP5 phosphorylation is up-regulated by interaction with NSP2.

Afrikanova

Fabbretti

Miozzo

et al. 1998

Journal of General Virology

129

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We have previously shown that a number of isoforms of the non-structural rotavirus protein NSP5 are found in virus-infected cells. These isoforms differ in their level of phosphorylation which, at least in part, appears to occur through autophosphorylation. NSP5 co-localizes with another non-structural protein, NSP2, in the viroplasms of infected cells where virus replication takes place. We now show that NSP5 can be chemically cross-linked in living cells with the viral polymerase VP1 and NSP2. Interaction of NSP5 with NSP2 was also demonstrated by coimmunoprecipitation of NSP2 and NSP5 from extracts of UV-treated rotavirus-infected cells. In

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Section: Introductionsupporting

confidence: 79%

Section: In Vivo Chemical Cross-linkingmentioning

confidence: 97%

Section: Immunoprecipitation and Western Immunoblot Analysismentioning

confidence: 99%

Section: Radiolabelling With [ 35 S]methionine Immunoprecipitationmentioning

confidence: 99%

Section: Radiolabelling With [ 35 S]methionine Immunoprecipitationmentioning

confidence: 99%

See 3 more Smart Citations

Rotavirus NSP5 phosphorylation is up-regulated by interaction with NSP2.

Afrikanova

Fabbretti

Miozzo

et al. 1998

Journal of General Virology

129

View full text Add to dashboard Cite

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Isolation and molecular characterization of a naturally occurring non-structural protein 5 (NSP5) gene reassortant of group A rotavirus of serotype G2P[4] with a long RNA pattern

Ahmed

Nakagomi

2005

J. Med. Virol.

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We report here two unusual strains of group A rotavirus, AU85 and AU102, isolated from children with diarrhea. These strains showed an unusual combination of serotype G2 and a long RNA pattern. RNA-RNA hybridization assays showed that these strains are reassortants in which a single genome segment 11 (the NSP5 gene) was derived from a Wa genogroup strain, while other 10 genome segments from a DS-1 genogroup strain. Phylogenetic analysis showed that the NSP5 gene of strain AU85 did not form cluster with Wa strain, while it belonged to the cluster of YM and other porcine strains. Phylogenetic analysis also showed that NSP5 and VP7 genes of AU85 were derived from the rotavirus circulating in the area. Both co-electrophoresis and RNA-RNA hybridization showed that AU85 and AU102 are identical strains. Moreover, the nucleotide sequence comparison between these two strains revealed that they had 100% identical NSP4, NSP5, and VP7 genes. These results suggest that AU85 was a reassortant formed relatively recently between rotaviruses belonging to the Wa and the DS-1 genogroup.

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Bluetongue Virus Assembly and Morphogenesis

Roy

Noad

Current Topics in Microbiology and Immunology

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Phosphorylation Generates Different Forms of Rotavirus NSP5

Cited by 72 publications

References 22 publications

Rotavirus NSP5 phosphorylation is up-regulated by interaction with NSP2.

Rotavirus NSP5 phosphorylation is up-regulated by interaction with NSP2.

Isolation and molecular characterization of a naturally occurring non-structural protein 5 (NSP5) gene reassortant of group A rotavirus of serotype G2P[4] with a long RNA pattern

Bluetongue Virus Assembly and Morphogenesis

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