Self-organization of the vesicular stomatitis virus nucleocapsid into a bullet shape

Desfosses, Ambroise; Ribeiro, Euripedes de Almeida; Schoehn, Guy; Blondel, Danielle; Guilligay, Delphine; Jamin, Marc; Ruigrok, Rob W.H.; Gutsche, Irina

doi:10.1038/ncomms2435

Cited by 27 publications

(27 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recent work has shown that N-RNA alone can adopt the classic bullet shape of the interior of the virion (43). This raises the possibility that the particles stuck at the membrane are viral particles that lack significant M. Earlier in vitro dissociation and reassociation experiments with NCM complexes proposed that the association of the RNP with a minor virion component mediates conversion to the NCM complex (12,13).…”

Section: Discussionmentioning

confidence: 99%

Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

Soh

Whelan

2015

J Virol

View full text Add to dashboard Cite

Vesicular stomatitis virus (VSV) assembly requires condensation of the viral ribonucleoprotein (RNP) core with the matrix protein (M) during budding from the plasma membrane. The RNP core comprises the negative-sense genomic RNA completely coated by the nucleocapsid protein (N) and associated by a phosphoprotein (P) with the large polymerase protein (L). To study the assembly of single viral particles, we tagged M and P with fluorescent proteins. We selected from a library of viruses with insertions in the M gene a replication-competent virus containing a fluorescent M and combined that with our previously described virus containing fluorescent P. Virus particles containing those fusions maintained the same bullet shape appearance as wild-type VSV but had a modest increase in particle length, reflecting the increased genome size. Imaging of the released particles revealed a variation in the amount of M and P assembled into the virions, consistent with a flexible packaging mechanism. We used the recombinants to further study the importance of the late domains in M, which serve to recruit the endosomal sorting complex required for transport (ESCRT) machinery during budding. Mutations in late domains resulted in the accumulation of virions that failed to pinch off from the plasma membrane. Imaging of single virions released from cells that were coinfected with M tagged with enhanced green fluorescent protein and M tagged with mCherry variants in which the late domains of one virus were inactivated by mutation showed a strong bias against the incorporation of the late-domain mutant into the released virions. In contrast, the intracellular expression and membrane association of the two variants were unaltered. These studies provide new tools for imaging particle assembly and enhance our resolution of existing models for assembly of VSV. IMPORTANCEAssembly of vesicular stomatitis virus (VSV) particles requires the separate trafficking of the viral replication machinery, a matrix protein (M) and a glycoprotein, to the plasma membrane. The matrix protein contains a motif termed a "late domain" that engages the host endosomal sorting complex required for transport (ESCRT) machinery to facilitate the release of viral particles. Inactivation of the late domains through mutation results in the accumulation of virions arrested at the point of release. In the study described here, we developed new tools to study VSV assembly by fusing fluorescent proteins to M and to a constituent of the replication machinery, the phosphoprotein (P). We used those tools to show that the late domains of M are required for efficient incorporation into viral particles and that the particles contain a variable quantity of M and P. V esicular stomatitis virus (VSV) forms particles with an ordered bullet shape. Those particles contain a lipid envelope modified by the insertion of the viral attachment and fusion glycoprotein (G). Inside the particle is a ribonucleoprotein (RNP) core that is delivered to the cytoplasm to initiate the process ...

show abstract

Section: Discussionmentioning

confidence: 99%

Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

Soh

Whelan

2015

J Virol

View full text Add to dashboard Cite

show abstract

“…In the virion, the N-RNA complex comprising 12 kb RNA and approximately 1300 N protomers is present in a highly condensed superhelical form and associated with approximately 650 copies of P protein dimers, which bind between two adjacent N molecules, and 50 molecules of the large RNA polymerase subunit L. The superhelix is surrounded by an envelope comprising a continuous inner M protein mesh, a lipid membrane, and trimers of the transmembrane glycoprotein G. The overall shape of the virion is imposed by the M protein layer, which determines the number of N subunits per turn, the diameter of the trunk, and the pitch of the RNP superhelix (Green et al, 2006). As described for VSV, the conical end is a consequence of N-RNA helix self-assembly and comprises the 5 -end of the viral RNP genome (Albertini et al, 2006;Desfosses et al, 2013;Green et al, 2006;Luo et al, 2007).…”

Section: Rabies Virionsmentioning

confidence: 99%

G gene-deficient single-round rabies viruses for neuronal circuit analysis

Ghanem

Conzelmann

2016

Virus Research

View full text Add to dashboard Cite

“…In vesicular stomatitis virus (VSV) of the Rhabdoviridae family, NC also forms a left-handed helix when packaged in the virion (19). When isolated from the virion, however, VSV NC does not retain a helical structure, although the vRNA is still sequestered in the nucleocapsid, according to the crystal structure of the VSV NLP (12,20). It has been shown that the C-terminal domain of VSV P (P CTD ) binds at the interface of two CTDs from neighboring parallel N subunits (21).…”

mentioning

confidence: 99%

Structural studies on the authentic mumps virus nucleocapsid showing uncoiling by the phosphoprotein

Cox

Pickar

Qiu

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Mumps virus (MuV) is a highly contagious pathogen, and despite extensive vaccination campaigns, outbreaks continue to occur worldwide. The virus has a negative-sense, single-stranded RNA genome that is encapsidated by the nucleocapsid protein (N) to form the nucleocapsid (NC). NC serves as the template for both transcription and replication. In this paper we solved an 18-Å-resolution structure of the authentic MuV NC using cryo-electron microscopy. We also observed the effects of phosphoprotein (P) binding on the MuV NC structure. The N-terminal domain of P (P NTD ) has been shown to bind NC and appeared to induce uncoiling of the helical NC. Additionally, we solved a 25-Å-resolution structure of the authentic MuV NC bound with the C-terminal domain of P (P CTD ). The location of the encapsidated viral genomic RNA was defined by modeling crystal structures of homologous negative strand RNA virus Ns in NC. Both the N-terminal and C-terminal domains of MuV P bind NC to participate in access to the genomic RNA by the viral RNA-dependent-RNA polymerase. These results provide critical insights on the structurefunction of the MuV NC and the structural alterations that occur through its interactions with P.replication | paramyxovirus | mononegavirale P aramyxoviruses are enveloped nonsegmented negative-strand RNA viruses (NSV) belonging to the order Mononegavirales. Mononegavirales also includes the Bornaviridae, Filoviridae, and Rhabdoviridae families. The Paramyxoviridae family includes several important human pathogens such as measles virus (MeV), respiratory syncytial virus (RSV), and mumps virus (MuV). Although vaccines exist for some paramyxoviruses, they are not available for others, such as RSV. In addition, no effective antiviral treatments have been developed.The MuV genome encodes 9 proteins, three of which are required for replication of the MuV genome; the nucleocapsid protein (N), phosphoprotein (P), and the large protein (L). N, P, and L have orthologs in a number of NSV. Studies on the roles of N, P, and L in viral RNA synthesis have shown that each can individually and differentially affect the processes of mRNA transcription and genome replication (1-10).Throughout the virus replication cycle, the genome of NSV always exists in the nucleocapsid (NC), a unique protein-RNA complex in which the viral RNA [viral genomic RNA (vRNA) or complementary genomic RNA (cRNA)] is completely sequestered by the N protein. NC is used as the functional template for RNA synthesis by the viral RNA dependent RNA polymerase (vRdRp), which includes L and P. The L protein contains all of the enzymatic activities needed for viral RNA synthesis, such as the ability to cap and polyadenylate mRNA transcripts. P acts as a cofactor to home vRdRp onto the NC template for RNA synthesis. In addition, the P protein chaperones monomeric and RNA-free N to encapsidate newly synthesized viral genomes during replication. The encapsidation of RNA by N is concomitant with the replication process.How the sequestered vRNA is accessed by vRdRp ...

show abstract

Self-organization of the vesicular stomatitis virus nucleocapsid into a bullet shape

Cited by 27 publications

References 29 publications

Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

G gene-deficient single-round rabies viruses for neuronal circuit analysis

Structural studies on the authentic mumps virus nucleocapsid showing uncoiling by the phosphoprotein

Contact Info

Product

Resources

About