Structure and protein composition of the rotavirus replicase particle

Patton, John T.; Gallegos, Claudia O.

doi:10.1016/0042-6822(88)90506-5

Cited by 58 publications

(56 citation statements)

References 25 publications

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“…NTPase activity of NSP2 is required for dsRNA synthesis. NSP2 has been implicated in rotavirus replication and packaging (6,27,29), but the contribution of the NTPase activity of NSP2 to these processes is uncertain. The enzymatic activity could be anticipated to have an essential function, given the conservation of the HIT-like motif in the NSP2a and NSP2c octamer and the retention of the activity.…”

Section: Vol 80 2006 Structure-function Analysis Of Rotavirus Nsp2 mentioning

confidence: 99%

Structure-Function Analysis of Rotavirus NSP2 Octamer by Using a Novel Complementation System

Taraporewala

Carpió

Jayaram

et al. 2006

J Virol

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Viral inclusion bodies, or viroplasms, that form in rotavirus-infected cells direct replication and packagingof the segmented double-stranded RNA (dsRNA) genome. NSP2, one of two rotavirus proteins needed for viroplasm assembly, possesses NTPase, RNA-binding, and helix-unwinding activities. NSP2 of the rotavirus group causing endemic infantile diarrhea (group A) was shown to self-assemble into large doughnut-shaped octamers with circumferential grooves and deep clefts containing nucleotide-binding histidine triad (HIT)-like motifs. Here, we demonstrate that NSP2 of group C rotavirus, a group that fails to reassort with group A viruses, retains the unique architecture of the group A octamer but differs in surface charge distribution. By using an NSP2-dependent complementation system, we show that the HIT-dependent NTPase activity of NSP2 is necessary for dsRNA synthesis, but not for viroplasm formation. The complementation system also showed that despite the retention of the octamer structure and the HIT-like fold, group C NSP2 failed to rescue replication and viroplasm formation in NSP2-deficient cells infected with group A rotavirus. The distinct differences in the surface charges on the Bristol and SA11 NSP2 octamers suggest that charge complementarity of the viroplasm-forming proteins guides the specificity of viroplasm formation and, possibly, reassortment restriction between rotavirus groups.

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Section: Vol 80 2006 Structure-function Analysis Of Rotavirus Nsp2 mentioning

confidence: 99%

Structure-Function Analysis of Rotavirus NSP2 Octamer by Using a Novel Complementation System

Taraporewala

Carpió

Jayaram

et al. 2006

J Virol

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“…Infective particles replicate in the cytoplasm of infected cells. Although several reports have described the characterization of rotavirus replication intermediates (Gallegos & Patton, 1989 ;Patton & Gallegos, 1988), molecular details of the replication mechanism remain unclear. Partially purified intermediate complexes (or sub-viral particles) obtained from SA11-infected cells were able to catalyse run-off synthesis of endogenous dsRNAs (Patton, 1986), as well as complete replication of exogenously added viral ss (j)-RNAs (Chen et al, 1994 ;Wentz et al, 1996 ;Patton et al, 1996 ;E.…”

Section: Introductionmentioning

confidence: 99%

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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“…These results suggest that viral transcription is catalysed by a multienzyme complex, present in the single-shelled particle, which requires enzymatic activities other than the viral RNA polymerase VP1 (Sandino et al, 1986;Valenzuela et aL, 1991). Viral RNA replication has also been found to occur in precore particles containing polypeptides VP 1, VP2 and VP3 and some of the non-structural proteins (Patton & Gallegos, 1988).…”

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confidence: 99%

“…The function of the viral polypeptides during viral RNA replication and transcription has been studied recently (Sandino et al, 1988;Patton & Gallegos, 1988Mansell & Patton, 1990;Valenzuela et al, 1991). Studies on in vitro transcription have allowed the identification of VP1 as the viral RNA polymerase (Valenzuela et al, 1991), VP3 as the guanylyltransferase, and VP2 as a single-and double-stranded RNA-binding protein Liu et al, 1992;Boyle & Holmes, 1986).…”

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Function of Rotavirus VP3 Polypeptide in Viral Morphogenesis

Vásquez¹,

Sandino²,

Pizarro³

et al. 1993

Journal of General Virology

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The phenotype of the rotavirus SA-11 mutant tsB carrying a thermosensitive mutation in gene 3, which encodes VP3, was characterized further from both infected cells and purified viral particles. The mutant phenotype was initially identified as negative for in v&o double-and single-stranded RNA synthesis. Our results show that the in vitro transcriptional properties of the tsB mutant at the restrictive temperature were identical to those of the wild-type strain. Similar results were obtained with respect to the VP3-associated guanylyltransferase activity. Analysis of viral particles made by mutant-infected cells at the restrictive temperature showed that only empty single-shelled particles were assembled. This indicates that viral morphogenesis is halted after the initial viral transcription and before RNA replication, suggesting that VP3 may be required as part of the replicase system but not for subviral particle assembly. These data suggest that such a phenotype is not due to alteration of a VP3 function related to transcription.

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Structure and protein composition of the rotavirus replicase particle

Cited by 58 publications

References 25 publications

Structure-Function Analysis of Rotavirus NSP2 Octamer by Using a Novel Complementation System

Structure-Function Analysis of Rotavirus NSP2 Octamer by Using a Novel Complementation System

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

Function of Rotavirus VP3 Polypeptide in Viral Morphogenesis

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