title: Goatpoxvirus ATPase activity is increased by dsDNA and decreased by zinc ion.
abstract: Viral-encoded ATPase can act as a part of molecular motor in genome packaging of DNA viruses, such as vaccinia virus and adenovirus, by ATP hydrolysis and interaction with DNA. Poxviral ATPase (also called A32) is involved in genomic double-stranded DNA (dsDNA) encapsidation, and inhibition of the expression of A32 causes formation of immature virions lacking viral DNA. However, the role of A32 in goatpoxvirus genome packaging and its dsDNA binding property are not known. In this study, purified recombinant goatpoxvirus A32 protein (rA32) was examined for its dsDNA binding property as well as the effect of dsDNA on ATP hydrolysis. We found that rA32 could bind dsDNA, and its ATPase activity was significant increased with dsDNA binding. Effects of magnesium and calcium ions on ATP hydrolysis were investigated also. The ATPase activity was dramatically enhanced by dsDNA in the presence of Mg(2+); in contrast, ATPase function was not altered by Ca(2+). Furthermore, the enzyme activity of rA32 was completely blocked by Zn(2+). Regarding DNA-protein interaction, the rA32-ATP-Mg(2+) showed lower dsDNA binding affinity than that of rA32-ATP-Ca(2+). The DNA-protein binding was stronger in the presence of zinc ion. Our results implied that A32 may play a role in viral genome encapsidation and DNA condensation.
authors.affiliation: Department of Veterinary Medicine, National Chiayi University, Chiayi City, 60061, Taiwan.
authors.forename: Ming-Liang
authors.initials: ML
authors.lastname: Lee
authors.affiliation: Graduate Institute of Veterinary Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung, 40227, Taiwan.
authors.forename: Wei-Li
authors.initials: WL
authors.lastname: Hsu
authors.affiliation: Department of Veterinary Medicine, College of Veterinary Medicine, National Chung-Hsing University, Taichung, 40227, Taiwan.
authors.forename: Chi-Young
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Department of Veterinary Medicine, National Chiayi University, Chiayi City, 60061, Taiwan.
authors.forename: Hui-Yu
authors.initials: HY
authors.lastname: Chen
authors.affiliation: Department of Veterinary Medicine, College of Veterinary Medicine, National Chung-Hsing University, Taichung, 40227, Taiwan.
authors.forename: Fong-Yuan
authors.initials: FY
authors.lastname: Lin
authors.affiliation: Department of Veterinary Medicine, National Chiayi University, Chiayi City, 60061, Taiwan.
authors.forename: Ming-Huang
authors.initials: MH
authors.lastname: Chang
authors.affiliation: Yunlin County Government, Yunlin County, 64001, Taiwan.
authors.forename: Hong-You
authors.initials: HY
authors.lastname: Chang
authors.affiliation: Department of Veterinary Medicine, College of Veterinary Medicine, National Chung-Hsing University, Taichung, 40227, Taiwan.
authors.forename: Min-Liang
authors.initials: ML
authors.lastname: Wong
authors.affiliation: Department of Veterinary Medicine, National Chiayi University, Chiayi City, 60061, Taiwan. luckyvet81@hotmail.com.
authors.forename: Kun-Wei
authors.initials: KW
authors.lastname: Chan
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D018151:Capripoxvirus / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000235:genetics*; D042003:DNA Packaging / Q000235:genetics; D004267:DNA Viruses / Q000235:genetics*; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D016679:Genome, Viral / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly / Q000235:genetics; D015032:Zinc / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D004247:DNA; D000251:Adenosine Triphosphatases; D015032:Zinc; D002118:Calcium
keywords: ATPase; DNA binding; DNA packaging; Goatpoxvirus; Zinc ion binding