Document Detail

A C-terminal, cysteine-rich site in poliovirus 2CATPase is required for morphogenesis.
MedLine Citation:
PMID:  24558221     Owner:  NLM     Status:  Publisher    
Morphogenesis of member viruses of genus Enterovirus of Picornaviridae is still poorly understood despite decades-long investigations. However, we recently provided evidence that 2C(ATPase) gives specificity to poliovirus encapsidation through an interaction with capsid protein VP3. Polypeptide 2C(ATPase) is a highly conserved nonstructural protein of enteroviruses with important roles in RNA replication, encapsidation and uncoating. We have identified a site (K279/R280) near the C-terminus of the polypeptide that is required for morphogenesis. The aim of our current project was to search for additional functional sites near the C-terminus of the 2C(ATPase) polypeptide, with particular interest in those that are required for encapsidation. We selected for analysis a cysteine-rich site of the polypeptide and constructed four mutants in which cysteines or a histidine was changed to an alanine. The RNA transcripts were transfected into HeLa cells yielding 2 lethal, one temperature-sensitive and one quasi-infectious mutant. All four mutants exhibited normal protein translation in vitro and 3 of them possessed severe RNA replication defects. The quasi-infectious mutant (C286A) yielded variants with a pseudoreversion at the original site (A286D) but some also contained one additional mutation: A138Vor M293V. The temperature-sensitive mutant (C272A/H273A) exhibited an encapsidation, and possibly also an uncoating defect, at 37°C. Variants of this mutant revealed suppressor mutations at 3 different sites in the 2C(ATPase) polypeptide: A138V, M293V, or K295R. We conclude that the cysteine-rich site near the C-terminus of 2C(ATPase) is involved in encapsidation, possibly through an interaction with an upstream segment located between boxes A and B of the nucleotide-binding domain.
Chunling Wang; Hsin-Chieh Ma; Eckard Wimmer; Ping Jiang; Aniko V Paul
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-2-20
Journal Detail:
Title:  The Journal of general virology     Volume:  -     ISSN:  1465-2099     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  2014 Feb 
Date Detail:
Created Date:  2014-2-21     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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