Time course of virus-specific macromolecular synthesis during rubella virus infection in vero cells

Hemphill, Mark L.; Forng, Ren-Yo; Abernathy, Emily; Frey, Teryl K.

doi:10.1016/0042-6822(88)90395-9

Cited by 65 publications

(65 citation statements)

References 39 publications

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“…The reason for the slow rate of entry is unknown but could involve slow migration of the virus receptor complex to a clathrincoated pit. RV infection is characterized by a long latent period (at least 12 h) and by the inability to achieve infection of all the cells at the time of initial virus adsorption (Hemphill et al, 1988). The slow kinetics of RV entry could be a significant factor in determining these characteristics.…”

Section: Discussionmentioning

confidence: 99%

Pathway of rubella virus infectious entry into Vero cells

Petruzziello

Orsi

Macchia

et al. 1996

Journal of General Virology

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The mechanism and the kinetics of rubella virus (RV) penetration into Vero cells were studied. By using pronase or acid treatment to inactivate virus which had adsorbed to cell membrane but had not been internalized, it was found that a period of 7 h was required in order for all of the adsorbed virus to enter the host cells. Lysosomotropic agents (monensin, methylamine, ammonium chloride and chloroquine) were used to study the mechanism by which RV penetrates host cells. Virus replication was inhibited if treatment of cells with these compounds was performed for at least 9 h after infection. However, if extracellular adsorbed virions were eliminated by acid treatment following removal of the lysosomotropic compounds, RV replication was completely inhibited by treatment with these drugs for any time period after adsorption. This indicated that the prolonged period of treatment with these compounds necessary to inhibit virus replication is due to the slow rate of RV internalization. None of the compounds had any effect on infection initiated by transfection of RV RNA, confirming that these drugs were exerting their inhibitory activity at penetration. The inhibition of RV replication by lysosomotropic compounds indicates that RV penetrates host cells by the endosomal pathway.

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Section: Discussionmentioning

confidence: 99%

Pathway of rubella virus infectious entry into Vero cells

Petruzziello

Orsi

Macchia

et al. 1996

Journal of General Virology

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“…RUB F-therien, BRD II, and RA27/3 strains were used in this study. The plaque assay was done as previously described (11). The protocol for reverse plaque assay, which is basically determining the number of puromycin resistant cells following transfection with a replicon which expresses puromycin acetyltransferase (PAC), has been described previously (4).…”

Section: Methodsmentioning

confidence: 99%

Rubella Virus Capsid Protein Modulates Viral Genome Replication and Virus Infectivity

Chen

Icenogle²

2004

J Virol

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The structural proteins (SP) of the Togaviridae can be deleted in defective interfering RNAs. The dispensability of viral SP has allowed construction of noninfectious viral expression vectors and replicons from viruses of the Alphavirus and Rubivirus genera. Nevertheless, in this study, we found that the SP of rubella virus (RUB) could enhance expression of reporter genes from RUB replicons in trans. SP enhancement required capsid protein (CP) expression and was not due to RNA-RNA recombination. Accumulation of minus-and plus-strand RNAs from replicons was observed in the presence of SP, suggesting that SP specifically affects RNA synthesis. By using replicons containing an antibiotic resistance gene, we found 2-to 50-fold increases in the number of cells surviving selection in the presence of SP. The increases depended significantly on the amount of transfected RNA. Small amounts of RNA or templates that replicated inefficiently showed more enhancement. The infectivity of infectious RNA was increased by at least 10-fold in cells expressing CP. Moreover, virus infectivity was greatly enhanced in such cells. In other cells that expressed higher levels of CP, RNA replication of replicons was inhibited. Thus, depending on conditions, CP can markedly enhance or inhibit RUB RNA replication.Rubella virus (RUB) is the sole member of Rubivirus genus in the Togaviridae family. RUB has a single-stranded, positivesense RNA genome that is 9,762 nucleotides (nt) in length. The genome encodes two open reading frames (ORFs): the 5Ј-proximal ORF encodes viral nonstructural proteins (NSP) that are responsible for viral genome replication, while the 3Ј-proximal ORF encodes three virion structural proteins (SP), the capsid protein (CP), and the two envelope glycoproteins, E2 and E1.A polyprotein, P200, is translated from the 5Ј-proximal ORF and is a major component of the replication complex for the synthesis of genomic minus-strand RNA, which serves as a template for the synthesis of plus-strand genomic and subgenomic RNA, the mRNA for the virion proteins. The switch of synthesis from minus-strand genomic RNAs to plus-strand RNA synthesis is dependent on the processing of P200 by the virally encoded protease (17), a mechanism similar to that observed in the NSP of Sindbis virus (16). Studies of RUB defective interfering (DI) RNAs have shown that DI RNAs generated during undiluted passaging of virus or persistently infected cells have most of the SP coding region removed, showing that most of the SP coding region is not required for replication of the viral genome (5, 6). RUB replicons have been constructed which have part of the SP coding region replaced by genes encoding the puromycin-resistance protein (PAC) (4) or green fluorescent protein (GFP) (28), respectively. Using the RUBrep/PAC, Chen et al. (4) identified the minimal region at the 3Ј end of the RUB genome that is required for viral genome replication and measured the replication efficiency of various 3Ј constructs. Using RUBrep/GFP, Tzeng et al. (28) showed that most o...

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“…In contrast, capsid is synthesized from a subgenomic mRNA and is a later event in the replication scheme (44). Typically, nonstructural and structural proteins are first detected in Vero cells at 12 and 16 h postinfection, respectively (19). Since the 6RA capsid does not bind well to genomic RNA, we used the 5RA strain for these experiments.…”

Section: 21 30)mentioning

confidence: 99%

Interactions between Rubella Virus Capsid and Host Protein p32 Are Important for Virus Replication

Beatch

Everitt²,

Law³

et al. 2005

J Virol

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The distribution and morphology of mitochondria are dramatically affected during infection with rubella virus (RV). Expression of the capsid, in the absence of other viral proteins, was found to induce both perinuclear clustering of mitochondria and the formation of electron-dense intermitochondrial plaques, both hallmarks of RV-infected cells. We previously identified p32, a host cell mitochondrial matrix protein, as a capsid-binding protein. Here, we show that two clusters of arginine residues within capsid are required for stable binding to p32. Mutagenic ablation of the p32-binding site in capsid resulted in decreased mitochondrial clustering, indicating that interactions with this cellular protein are required for capsid-dependent reorganization of mitochondria. Recombinant viruses encoding arginine-to-alanine mutations in the p32-binding region of capsid exhibited altered plaque morphology and replicated to lower titers. Further analysis indicated that disruption of stable interactions between capsid and p32 was associated with decreased production of subgenomic RNA and, consequently, infected cells produced significantly lower amounts of viral structural proteins under these conditions. Together, these results suggest that capsid-p32 interactions are important for nonstructural functions of capsid that include regulation of virus RNA replication and reorganization of mitochondria during infection.

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Time course of virus-specific macromolecular synthesis during rubella virus infection in vero cells

Cited by 65 publications

References 39 publications

Pathway of rubella virus infectious entry into Vero cells

Pathway of rubella virus infectious entry into Vero cells

Rubella Virus Capsid Protein Modulates Viral Genome Replication and Virus Infectivity

Interactions between Rubella Virus Capsid and Host Protein p32 Are Important for Virus Replication

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