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PMID: 20865167 Published · epublish English Journal Article Research Support, N.I.H., Extramural

Direct interaction between two viral proteins, the nonstructural protein 2C and the capsid protein VP3, is required for enterovirus morphogenesis.

PLoS pathogens ·Vol. 6 ·No. 8 ·2010-08-26 ·Pages e1001066

Liu Y, Wang C, Mueller S, Paul AV, Wimmer E, Jiang P

Abstract

In spite of decades-long studies, the mechanism of morphogenesis of plus-stranded RNA viruses belonging to the genus Enterovirus of Picornaviridae, including poliovirus (PV), is not understood. Numerous attempts to identify an RNA encapsidation signal have failed. Genetic studies, however, have implicated a role of the non-structural protein 2C(ATPase) in the formation of poliovirus particles. Here we report a novel mechanism in which protein-protein interaction is sufficient to explain the specificity in PV encapsidation. Making use of a novel "reporter virus", we show that a quasi-infectious chimera consisting of the capsid precursor of C-cluster coxsackie virus 20 (C-CAV20) and the nonstructural proteins of the closely related PV translated and replicated its genome with wild type kinetics, whereas encapsidation was blocked. On blind passages, encapsidation of the chimera was rescued by a single mutation either in capsid protein VP3 of CAV20 or in 2C(ATPase) of PV. Whereas each of the single-mutation variants expressed severe proliferation phenotypes, engineering both mutations into the chimera yielded a virus encapsidating with wild type kinetics. Biochemical analyses provided strong evidence for a direct interaction between 2C(ATPase) and VP3 of PV and CAV20. Chimeras of other C-CAVs (CAV20/CAV21 or CAV18/CAV20) were blocked in encapsidation (no virus after blind passages) but could be rescued if the capsid and 2C(ATPase) coding regions originated from the same virus. Our novel mechanism explains the specificity of encapsidation without apparent involvement of an RNA signal by considering that (i) genome replication is known to be stringently linked to translation, (ii) morphogenesis is known to be stringently linked to genome replication, (iii) newly synthesized 2C(ATPase) is an essential component of the replication complex, and (iv) 2C(ATPase) has specific affinity to capsid protein(s). These conditions lead to morphogenesis at the site where newly synthesized genomes emerge from the replication complex.

MeSH Terms
Capsid Proteins/genetics,metabolism Carrier Proteins/genetics,metabolism Enterovirus/physiology HeLa Cells Humans Immunoprecipitation Poliovirus/genetics,metabolism RNA, Viral/genetics Reverse Transcriptase Polymerase Chain Reaction Viral Nonstructural Proteins/genetics,metabolism Viral Proteins/genetics,metabolism Virus Replication/physiology
Chemicals
Capsid Proteins Carrier Proteins RNA, Viral VP3 protein, Poliovirus Viral Nonstructural Proteins Viral Proteins 2C protein, viral
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Liu Ying
Department of Molecular Genetics and Microbiology, School of Medicine, Stony Brook University, Stony Brook, New York, United States of America.
Wang Chunling
Mueller Steffen
Paul Aniko V
Wimmer Eckard
Jiang Ping
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Article Info
Journal
PLoS pathogens
Abbr.
PLoS Pathog
ISSN
1553-7374
DOI
10.1371/journal.ppat.1001066
Published
2010-08-26
Epub
2010-00-26
Pages
e1001066
Language
English
Region
United States
NLM ID
101238921
PMCID
PMC2928791
Subset
IM
Grants
NIAID NIH HHS · R01 AI015122 · United States
NIAID NIH HHS · R37 AI015122 · United States
NIAID NIH HHS · AI 15122 · United States
External Links
PubMed source DOI: 10.1371/journal.ppat.1001066 PMC Full Text View journal details
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