Respiration and ATP Level in BHK21/13S Cells during the Earliest Stages of Rubella Virus Replication

Bardeletti, Gilbert

doi:10.1159/000148884

Cited by 25 publications

(15 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mitochondria have long been thought to have an important role in the replication of RV (1,2,33). Moreover, we recently demonstrated that expression of the RV structural proteins induces relocalization of mitochondria to the perinuclear region (6).…”

Section: Resultsmentioning

confidence: 99%

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

Beatch

Everitt²,

Law³

et al. 2005

J Virol

View full text Add to dashboard Cite

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.

show abstract

Section: Resultsmentioning

confidence: 99%

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

Beatch

Everitt²,

Law³

et al. 2005

J Virol

View full text Add to dashboard Cite

show abstract

“…The distribution of ribosomes in RV-infected cells thus appeared to be unaffected, as observed in alphavirus-infected cells (6) but distinct from the profound changes reported in flavivirus-infected cells (15). Similarly, with mitochondrial antibodies, no change in the distribution of mitochondria was noted between mock-infected and infected cells at 24 hours, providing no support for the suggestion that the mitochondria may be involved in RV replication and maturation (2).…”

Section: Distribution Of Rubella Virus Antigens In Infected Cellsmentioning

confidence: 84%

Distribution by immunofluorescence of viral products and actin-containing cytoskeletal filaments in rubella virus-infected cells

Bowden

Pedersen

Toh

et al. 1987

Archives of Virology

View full text Add to dashboard Cite

Rubella virus (RV)-host cell interactions were examined by indirect immunofluorescence staining using antibodies to viral products and cytoskeletal components as probes. The patterns of immunofluorescence observed with human convalescent sera indicated that in infected Vero cells RV-specified proteins were distributed throughout the rough endoplasmic reticulum with some possible accumulation in the region of the Golgi complex. Viral RNA synthesis, detected with anti-double stranded RNA, appeared to be confined to small, intensely stained foci irregularly distributed in the cytoplasm. When cells were infected at a higher multiplicity, these foci appeared to aggregate into linear arrays. Infection with RV had a profound effect on the organization of actin in both Vero and BHK 21 cells, as shown by anti-actin antibodies. Actin microfilaments were observed to disintegrate progressively into amorphous aggregates of apparently monomeric actin as the infection proceeded. Because of the role actin microfilaments may play in cell mitosis it is postulated that this effect may be related to the inhibition of cell division reported to be associated with the congenital rubella syndrome.

show abstract

“…The localization of capsid to the mitochondria and association with a mitochondrial protein is intriguing in light of previous studies which document a link between RV replication complexes and mitochondrial function and localization (2,3,24,25). In RV-infected cells, clustering of mitochondria occurs and electron-dense plaques form between the membranes of apposing mitochondria and between mitochondria that associate with the endoplasmic reticulum.…”

Section: Discussionmentioning

confidence: 96%

Rubella Virus Capsid Associates with Host Cell Protein p32 and Localizes to Mitochondria

Beatch

Hobman

2000

J Virol

View full text Add to dashboard Cite

Togavirus nucleocapsids have a characteristic icosahedral structure and are composed of multiple copies of a capsid protein complexed with genomic RNA. The assembly of rubella virus nucleocapsids is unique among togaviruses in that the process occurs late in virus assembly and in association with intracellular membranes. The goal of this study was to identify host cell proteins which may be involved in regulating rubella virus nucleocapsid assembly through their interactions with the capsid protein. Capsid was used as bait to screen a CV1 cDNA library using the yeast two-hybrid system. One protein that interacted strongly with capsid was p32, a cellular protein which is known to interact with other viral proteins. The interaction between capsid and p32 was confirmed using a number of different in vitro and in vivo methods, and the site of interaction between these two proteins was shown to be at the mitochondria. Interestingly, overexpression of the rubella virus structural proteins resulted in clustering of the mitochondria in the perinuclear region. The p32-binding site in capsid is a potentially phosphorylated region that overlaps the viral RNA-binding domain of capsid. Our results are consistent with the possibility that the interaction of p32 with capsid plays a role in the regulation of nucleocapsid assembly and/or virus-host interactions.Rubella virus (RV) is a positive-strand RNA virus of the family Togaviridae. Despite routine vaccination programs which have been in place for 30 years, the virus persists in the human population and remains an important human pathogen (11). The most serious medical consequences of RV infection occur when seronegative women contract the virus during the first trimester of pregnancy. RV is highly teratogenic and causes a characteristic pattern of defects in the fetus which are collectively known as congenital rubella syndrome. The molecular basis of RV pathology remains poorly understood, but several recent reports have shown that the virus induces apoptosis in a cell type-dependent manner (9,19,33,45). This pattern of apoptosis could potentially explain the organ-specific malformations observed in congenital rubella syndrome.Virions contain three structural proteins that are translated from a 24S subgenomic RNA; two membrane-spanning glycoproteins (E2 and E1) and a capsid protein (38). The capsid protein is multifunctional and is involved in several different types of intermolecular interactions. First, it contains an RNAbinding domain and is responsible for packaging the genomic RNA into nucleocapsids (11,28). Second, by analogy with other togaviruses, capsid must engage in homo-oligomeric (capsid-capsid) interactions during nucleocapsid formation. Finally, it must also interact with E2 and/or E1 during budding (37, 40).The nucleocapsids of togaviruses have a characteristic icosahedral structure which has been extensively studied in alphaviruses (41). Although the overall structures of RV and alphavirus capsids are similar, their assembly pathways are quite different. Wh...

show abstract

Respiration and ATP Level in BHK21/13S Cells during the Earliest Stages of Rubella Virus Replication

Cited by 25 publications

References 13 publications

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

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

Distribution by immunofluorescence of viral products and actin-containing cytoskeletal filaments in rubella virus-infected cells

Rubella Virus Capsid Associates with Host Cell Protein p32 and Localizes to Mitochondria

Contact Info

Product

Resources

About