title: Periplaneta fuliginosa densovirus nonstructural protein NS1 contains an endonuclease activity that is regulated by its phosphorylation.
abstract: Periplaneta fuliginosa densovirus (PfDNV) is a single-stranded DNA virus, belonging to Densovirinae subfamily, Parvoviridae family. Parvovirus nonstructural protein 1 (NS1) contains various activities required for parvoviral DNA replication, like endonuclease, helicase and ATPase, which are regulated by serine/threonine phosphorylation. However, for PfDNV, NS1 endonuclease activity has not been determined. Moreover, for densoviruses, whether NS1 is phosphorylated, and if so, phosphorylation pattern and impact on NS1 activities have not been investigated. Here, we demonstrated that PfDNV NS1 possesses endonuclease activity, covalently attaches to 5'-end of nicking site, and includes an active-site tyrosine (Y178). Moreover, using different phosphatases, we uncovered that both serine/threonine and tyrosine phosphorylations are critical for NS1 endonuclease and helicase activities. Further mass-spec and mutational analyses revealed that Y345 is phosphorylated and functions as a critical regulatory site for NS1 activities. This study should foster our understanding of NS1 activities and regulations in PfDNV and other densoviruses.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
authors.forename: Yajuan
authors.initials: Y
authors.lastname: Han
authors.forename: Qinrong
authors.initials: Q
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Qiu
authors.forename: Wenzhe
authors.initials: W
authors.lastname: Wu
authors.forename: Huihui
authors.initials: H
authors.lastname: He
authors.forename: Jiamin
authors.initials: J
authors.lastname: Zhang
authors.forename: Yuanyang
authors.initials: Y
authors.lastname: Hu
authors.forename: Xi
authors.initials: X
authors.lastname: Zhou
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D004265:DNA Helicases / Q000378:metabolism*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000378:metabolism; D017990:Densovirus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004720:Endonucleases / Q000378:metabolism*; D009154:Mutation; D010526:Periplaneta / Q000821:virology*; D010766:Phosphorylation; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D017361:Viral Nonstructural Proteins; D004720:Endonucleases; D004265:DNA Helicases