title: Putative phosphorylation sites on WCA domain of HA2 is essential for Helicoverpa armigera single nucleopolyhedrovirus replication.
abstract: Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality. The Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain, in which phosphorylation status are supposed to be critical in respect to actin polymerization. In the present study, two putative phosphorylation sites ((232)Thr and (250)Ser) and a highly conserved Serine ((245)Ser) on the WCA domain of HA2 were mutated, and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome. Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at (245)Ser can produce infectious virions, both (232)Thr and (250)Ser mutations were lethal to the virus. However, actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus, which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway.
authors.affiliation: Key Laboratory of Agricultural and Enviromental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Yi-pin
authors.initials: YP
authors.lastname: Lv
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Chun-chen
authors.initials: CC
authors.lastname: Wu
authors.forename: Rong-juan
authors.initials: RJ
authors.lastname: Pei
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
authors.forename: Xin-wen
authors.initials: XW
authors.lastname: Chen
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D019295:Computational Biology; D004261:DNA Replication / Q000235:genetics / Q000502:physiology*; D010766:Phosphorylation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics / Q000378:metabolism*; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; C526765:HA2 protein, Helicoverpa armigera nucleopolyhedrovirus; D014764:Viral Proteins; D051316:Wiskott-Aldrich Syndrome Protein; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal