scholarly journals Analyses of Phosphorylation Events in the Rubella Virus Capsid Protein: Role in Early Replication Events

2006 ◽  
Vol 80 (14) ◽  
pp. 6917-6925 ◽  
Author(s):  
LokMan J. Law ◽  
Carolina S. Ilkow ◽  
Wen-Pin Tzeng ◽  
Matthew Rawluk ◽  
David T. Stuart ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Rubella Virus ◽  
Rna Binding ◽  
Virus Assembly ◽  
Binding Activity ◽  
Amino Acid Residues ◽  
Direct Role ◽  
Virus Capsid ◽  
Early Replication ◽  
Virus Capsid Protein

ABSTRACT The Rubella virus capsid protein is phosphorylated prior to virus assembly. Our previous data are consistent with a model in which dynamic phosphorylation of the capsid regulates its RNA binding activity and, in turn, nucleocapsid assembly. In the present study, the process of capsid phosphorylation was examined in further detail. We show that phosphorylation of serine 46 in the RNA binding region of the capsid is required to trigger phosphorylation of additional amino acid residues that include threonine 47. This residue likely plays a direct role in regulating the binding of genomic RNA to the capsid. We also provide evidence which suggests that the capsid is dephosphorylated prior to or during virus budding. Finally, whereas the phosphorylation state of the capsid does not directly influence the rate of synthesis of viral RNA and proteins or the assembly and secretion of virions, the presence of phosphate on the capsid is critical for early events in virus replication, most likely the uncoating of virions and/or disassembly of nucleocapsids.

scholarly journals Rubella virus capsid protein structure and its role in virus assembly and infection

2013 ◽  
Vol 110 (50) ◽  
pp. 20105-20110 ◽  
Author(s):  
V. Mangala Prasad ◽  
S. D. Willows ◽  
A. Fokine ◽  
A. J. Battisti ◽  
S. Sun ◽  
...  
Keyword(s):  
Protein Structure ◽  
Capsid Protein ◽  
Rubella Virus ◽  
Virus Assembly ◽  
Virus Capsid ◽  
Virus Capsid Protein

scholarly journals Analysis of Rubella Virus Capsid Protein-Mediated Enhancement of Replicon Replication and Mutant Rescue

2006 ◽  
Vol 80 (8) ◽  
pp. 3966-3974 ◽  
Author(s):  
Wen-Pin Tzeng ◽  
Jason D. Matthews ◽  
Teryl K. Frey
Keyword(s):  
Capsid Protein ◽  
Rubella Virus ◽  
Fluorescent Protein ◽  
Vero Cells ◽  
Basic Step ◽  
Virus Capsid ◽  
Replicase Protein ◽  
Virus C ◽  
Green Fluorescent ◽  
Virus Capsid Protein

ABSTRACT The rubella virus capsid protein (C) has been shown to complement a lethal deletion (termed ΔNotI) in P150 replicase protein. To investigate this phenomenon, we generated two lines of Vero cells that stably expressed either C (C-Vero cells) or C lacking the eight N-terminal residues (CΔ8-Vero cells), a construct previously shown to be unable to complement ΔNotI. In C-Vero cells but not Vero or CΔ8-Vero cells, replication of a wild-type (wt) replicon expressing the green fluorescent protein (GFP) reporter gene (RUBrep/GFP) was enhanced, and replication of a replicon with ΔNotI (RUBrep/GFP-ΔNotI) was rescued. Surprisingly, replicons with deleterious mutations in the 5′ and 3′ cis-acting elements were also rescued in C-Vero cells. Interestingly, the CΔ8 construct localized to the nucleus while the C construct localized in the cytoplasm, explaining the lack of enhancement and rescue in CΔ8-Vero cells since rubella virus replication occurs in the cytoplasm. Enhancement and rescue in C-Vero cells were at a basic step in the replication cycle, resulting in a substantial increase in the accumulation of replicon-specific RNAs. There was no difference in translation of the nonstructural proteins in C-Vero and Vero cells transfected with the wt and mutant replicons, demonstrating that enhancement and rescue were not due to an increase in the efficiency of translation of the transfected replicon transcripts. In replicon-transfected C-Vero cells, C and the P150 replicase protein associated by coimmunoprecipitation, suggesting that C might play a role in RNA replication, which could explain the enhancement and rescue phenomena. A unifying model that accounts for enhancement of wt replicon replication and rescue of diverse mutations by the rubella virus C protein is proposed.

scholarly journals RNA Binding by the Brome Mosaic Virus Capsid Protein and the Regulation of Viral RNA Accumulation

2009 ◽  
Vol 391 (2) ◽  
pp. 314-326 ◽  
Author(s):  
Guanghui Yi ◽  
Robert C. Vaughan ◽  
Ian Yarbrough ◽  
S. Dharmaiah ◽  
C. Cheng Kao
Keyword(s):  
Mosaic Virus ◽  
Capsid Protein ◽  
Rna Binding ◽  
Brome Mosaic Virus ◽  
Viral Rna ◽  
Virus Capsid ◽  
Rna Accumulation ◽  
Virus Capsid Protein

scholarly journals Drug Repurposing Commonly Against Dengue Virus Capsid and SARS-CoV-2 Nucleocapsid: An in Silico Approach

2020 ◽  
Author(s):  
Debica Mukherjee ◽  
Rupesh Roy ◽  
UPASANA RAY
Keyword(s):  
Dengue Virus ◽  
Capsid Protein ◽  
In Silico ◽  
Rna Binding ◽  
Drug Repurposing ◽  
Virus Capsid ◽  
Approved Drugs ◽  
Full Swing ◽  
Fda Approved Drugs ◽  
Virus Capsid Protein

<p></p><p>In the middle of SARS-CoV-2 pandemic, dengue virus (DENV) is giving a silent warning as the season approaches nearer. There is no specific antiviral against DENV for use in the clinics. Thus, considering these facts we can potentially face both these viruses together increasing the clinical burden. The search for anti-viral drugs against SARS-CoV-2 is in full swing and repurposing of already ‘in-use’ drugs against other diseases or COVID-19 has drawn significant attention. Earlier we had reported few FDA approved anti-viral and anti-microbial drugs that could be tested for binding with SARS-CoV-2 nucleocapsid N terminal domain. We explored the possibility of interactions of the drugs screened for SARS-CoV2 with Dengue virus capsid protein. We report five FDA approved drugs that were seen to be docking onto the SARS-CoV-2 nucleocapsid RNA binding domain, also docking well with DENV capsid protein on the RNA binding site and/or the capsid’s membrane fusion domain. Thus, the present study proposes these five drugs as common antiviral candidates against both SARS-CoV-2 and DENV although the <i>in silico</i> study is subject to further validations.</p><br><p></p>

scholarly journals Rubella Virus Capsid Protein Induces Apoptosis in Transfected RK13 Cells

Virology ◽  
2000 ◽  
Vol 275 (1) ◽  
pp. 20-29 ◽  
Author(s):  
Robert Duncan ◽  
Ali Esmaili ◽  
Lok Man J. Law ◽  
Sylvie Bertholet ◽  
Chris Hough ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Rubella Virus ◽  
Virus Capsid ◽  
Virus Capsid Protein

scholarly journals Functional Requirements of the Yellow Fever Virus Capsid Protein

2007 ◽  
Vol 81 (12) ◽  
pp. 6471-6481 ◽  
Author(s):  
Chinmay G. Patkar ◽  
Christopher T. Jones ◽  
Yu-hsuan Chang ◽  
Ranjit Warrier ◽  
Richard J. Kuhn
Keyword(s):  
Capsid Protein ◽  
Yellow Fever ◽  
Yellow Fever Virus ◽  
Virus Assembly ◽  
Fever Virus ◽  
Site Directed Mutagenesis ◽  
Packaging System ◽  
Virus Capsid ◽  
Double Mutation ◽  
Virus Capsid Protein

ABSTRACT Although it is known that the flavivirus capsid protein is essential for genome packaging and formation of infectious particles, the minimal requirements of the dimeric capsid protein for virus assembly/disassembly have not been characterized. By use of a trans-packaging system that involved packaging a yellow fever virus (YFV) replicon into pseudo-infectious particles by supplying the YFV structural proteins using a Sindbis virus helper construct, the functional elements within the YFV capsid protein (YFC) were characterized. Various N- and C-terminal truncations, internal deletions, and point mutations of YFC were analyzed for their ability to package the YFV replicon. Consistent with previous reports on the tick-borne encephalitis virus capsid protein, YFC demonstrates remarkable functional flexibility. Nearly 40 residues of YFC could be removed from the N terminus while the ability to package replicon RNA was retained. Additionally, YFC containing a deletion of approximately 27 residues of the C terminus, including a complete deletion of C-terminal helix 4, was functional. Internal deletions encompassing the internal hydrophobic sequence in YFC were, in general, tolerated to a lesser extent. Site-directed mutagenesis of helix 4 residues predicted to be involved in intermonomeric interactions were also analyzed, and although single mutations did not affect packaging, a YFC with the double mutation of leucine 81 and valine 88 was nonfunctional. The effects of mutations in YFC on the viability of YFV infection were also analyzed, and these results were similar to those obtained using the replicon packaging system, thus underscoring the flexibility of YFC with respect to the requirements for its functioning.

Baculovirus-Mediated Large-Scale Expression and Purification of a Polyhistidine-Tagged Rubella Virus Capsid Protein

1998 ◽  
Vol 12 (3) ◽  
pp. 323-330 ◽  
Author(s):  
Michel Schmidt ◽  
Nina Tuominen ◽  
Tove Johansson ◽  
Stefan A. Weiss ◽  
Kari Keinänen ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Rubella Virus ◽  
Large Scale ◽  
Expression And Purification ◽  
Virus Capsid ◽  
Virus Capsid Protein

scholarly journals Phosphorylation of Rubella Virus Capsid Regulates Its RNA Binding Activity and Virus Replication

2003 ◽  
Vol 77 (3) ◽  
pp. 1764-1771 ◽  
Author(s):  
Lok Man J. Law ◽  
Jason C. Everitt ◽  
Martin D. Beatch ◽  
Charles F. B. Holmes ◽  
Tom C. Hobman
Keyword(s):  
Virus Replication ◽  
Rubella Virus ◽  
Rna Binding ◽  
Virus Assembly ◽  
Binding Activity ◽  
Viral Rna ◽  
Wild Type Virus ◽  
Genomic Rna ◽  
Wild Type ◽  
Positive Strand Rna

ABSTRACT Rubella virus is an enveloped positive-strand RNA virus of the family Togaviridae. Virions are composed of three structural proteins: a capsid and two membrane-spanning glycoproteins, E2 and E1. During virus assembly, the capsid interacts with genomic RNA to form nucleocapsids. In the present study, we have investigated the role of capsid phosphorylation in virus replication. We have identified a single serine residue within the RNA binding region that is required for normal phosphorylation of this protein. The importance of capsid phosphorylation in virus replication was demonstrated by the fact that recombinant viruses encoding hypophosphorylated capsids replicated at much lower titers and were less cytopathic than wild-type virus. Nonphosphorylated mutant capsid proteins exhibited higher affinities for viral RNA than wild-type phosphorylated capsids. Capsid protein isolated from wild-type strain virions bound viral RNA more efficiently than cell-associated capsid. However, the RNA-binding activity of cell-associated capsids increased dramatically after treatment with phosphatase, suggesting that the capsid is dephosphorylated during virus assembly. In vitro assays indicate that the capsid may be a substrate for protein phosphatase 1A. As capsid is heavily phosphorylated under conditions where virus assembly does not occur, we propose that phosphorylation serves to negatively regulate binding of viral genomic RNA. This may delay the initiation of nucleocapsid assembly until sufficient amounts of virus glycoproteins accumulate at the budding site and/or prevent nonspecific binding to cellular RNA when levels of genomic RNA are low. It follows that at a late stage in replication, the capsid may undergo dephosphorylation before nucleocapsid assembly occurs.

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