Mutational analysis of the herpes simplex virus triplex protein VP19C

Adamson, W.E., McNab, D., Preston, V.G. and Rixon, F.J. (2006) Mutational analysis of the herpes simplex virus triplex protein VP19C. Journal of Virology, 80(3), pp. 1537-1548. (doi: 10.1128/JVI.80.3.1537-1548.2006) (PMID:16415029) (PMCID:PMC1346951)

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Herpes simplex virus type 1 (HSV-1) capsids have an icosahedral structure with capsomers formed by the major capsid protein, VP5, linked in groups of three by distinctive structures called triplexes. Triplexes are heterotrimers formed by two proteins in a 1:2 stoichiometry. The single-copy protein is called VP19C, and the dimeric protein is VP23. We have carried out insertional and deletional mutagenesis on VP19C and have examined the effects of the mutations on virus growth and capsid assembly. Insertional mutagenesis showed that the N-terminal ~100 amino acids of the protein, which correspond to a region that is poorly conserved among herpesviruses, are insensitive to disruption and that insertions into the rest of the protein had various effects on virus growth. Some, but not all, severely disabled mutants were compromised in the ability to bind VP23 or VP5. Analysis of deletion mutants revealed the presence of a nuclear localization signal (NLS) near the N terminus of VP19C, and this was mapped to a 33-amino-acid region by fusion of specific sequences to a green fluorescent protein marker. By replacing the endogenous NLS with that from the simian virus 40 large T antigen, we were able to show that the first 45 amino acids of VP19C were not essential for assembly of functional capsids and infectious virus particles. However, removing the first 63 amino acids resulted in formation of aberrant capsids and prevented virus growth, suggesting that the poorly conserved N-terminal sequences have some as-yet-unidentified function.

Item Type:Articles
Glasgow Author(s) Enlighten ID:McNab, Mr David and Rixon, Dr Frazer and Preston, Dr Valerie
Authors: Adamson, W.E., McNab, D., Preston, V.G., and Rixon, F.J.
Subjects:Q Science > QR Microbiology > QR355 Virology
College/School:College of Medical Veterinary and Life Sciences
Journal Name:Journal of Virology
Journal Abbr.:J. Virol.
Publisher:American Society for Microbiology
ISSN (Online):1098-5514

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
656531Structural studies on human virusesFrazer RixonMedical Research Council (MRC)MC_UU_12014/6MVLS III - CENTRE FOR VIRUS RESEARCH