title: Mutational analysis of varicella-zoster virus (VZV) immediate early protein (IE62) subdomains and their importance in viral replication.
abstract: VZV IE62 is an essential, immediate-early, tegument protein and consists of five domains. We generated recombinant viruses carrying mutations in the first three IE62 domains and tested their influence on VZV replication kinetics. The mutations in domain I did not affect replication kinetics while domain II mutations, disrupting the DNA binding and dimerization domain (DBD), were lethal for VZV replication. Mutations in domain III of the nuclear localization signal (NLS) and the two phosphorylation sites S686A/S722A resulted in slower growth in early and late infection respectively and were associated with IE62 accumulation in the cytoplasm and nucleus respectively. This study mapped the functional domains of IE62 in context of viral infection, indicating that DNA binding and dimerization domain is essential for VZV replication. In addition, the correct localization of IE62, whether nuclear or cytoplasmic, at different points in the viral life cycle, is important for normal progression of VZV replication.
authors.affiliation: Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA, United States; Department of Molecular Biology, National Research Centre, El-Buhouth St., Cairo, Egypt. Electronic address: mkhalil2@stanford.edu.
authors.forename: Mohamed I
authors.initials: MI
authors.lastname: Khalil
authors.affiliation: Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA, United States.
authors.forename: Xibing
authors.initials: X
authors.lastname: Che
authors.affiliation: Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA, United States.
authors.forename: Phillip
authors.initials: P
authors.lastname: Sung
authors.affiliation: Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA, United States.
authors.forename: Marvin H
authors.initials: MH
authors.lastname: Sommer
authors.affiliation: Department of Microbiology and Immunology, School of Medicine and Biomedical Science, University at Buffalo, Buffalo, NY, United States.
authors.forename: John
authors.initials: J
authors.lastname: Hay
authors.affiliation: Departments of Pediatrics and Microbiology & Immunology, Stan ford University School of Medicine, Stanford, CA, United States.
authors.forename: Ann M
authors.initials: AM
authors.lastname: Arvin
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology; D015870:Gene Expression; D017930:Genes, Reporter; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D019913:Nuclear Localization Signals / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D016865:Reassortant Viruses / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
chemical_list: C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D019913:Nuclear Localization Signals; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D008156:Luciferases
keywords: DBD; Dimerization; IE62; NLS; Putative phosphorylation site; TAD; VZV