title: Parvovirus B19 NS1 protein induces cell cycle arrest at G2-phase by activating the ATR-CDC25C-CDK1 pathway.
abstract: Human parvovirus B19 (B19V) infection of primary human erythroid progenitor cells (EPCs) arrests infected cells at both late S-phase and G2-phase, which contain 4N DNA. B19V infection induces a DNA damage response (DDR) that facilitates viral DNA replication but is dispensable for cell cycle arrest at G2-phase; however, a putative C-terminal transactivation domain (TAD2) within NS1 is responsible for G2-phase arrest. To fully understand the mechanism underlying B19V NS1-induced G2-phase arrest, we established two doxycycline-inducible B19V-permissive UT7/Epo-S1 cell lines that express NS1 or NS1mTAD2, and examined the function of the TAD2 domain during G2-phase arrest. The results confirm that the NS1 TAD2 domain plays a pivotal role in NS1-induced G2-phase arrest. Mechanistically, NS1 transactivated cellular gene expression through the TAD2 domain, which was itself responsible for ATR (ataxia-telangiectasia mutated and Rad3-related) activation. Activated ATR phosphorylated CDC25C at serine 216, which in turn inactivated the cyclin B/CDK1 complex without affecting nuclear import of the complex. Importantly, we found that the ATR-CHK1-CDC25C-CDK1 pathway was activated during B19V infection of EPCs, and that ATR activation played an important role in B19V infection-induced G2-phase arrest.
authors.affiliation: School of Life Sciences, Central China Normal University, Wuhan, China.|Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.affiliation: Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing, China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Department of Pediatrics and Department of Biomedical and Health Informatics, The Children's Mercy Hospital and University of Missouri Kansas City School of Medicine, Kansas City, Missouri, United States of America.
authors.forename: Min
authors.initials: M
authors.lastname: Xiong
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Wei
authors.initials: W
authors.lastname: Zou
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Deng
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Safder S
authors.initials: SS
authors.lastname: Ganaie
authors.affiliation: Department of Research and Development, Viracor-IBT Laboratories, Lee's Summit, Missouri, United States of America.
authors.forename: Steve
authors.identifier: 0000-0002-4840-7710
authors.initials: S
authors.lastname: Kleiboeker
authors.affiliation: School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Peng
authors.affiliation: School of Life Sciences, Central China Normal University, Wuhan, China.
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Liu
authors.affiliation: Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing, China.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Pediatrics and Department of Biomedical and Health Informatics, The Children's Mercy Hospital and University of Missouri Kansas City School of Medicine, Kansas City, Missouri, United States of America.
authors.forename: Shui Qing
authors.initials: SQ
authors.lastname: Ye
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Jianming
authors.identifier: 0000-0001-9850-1695
authors.initials: J
authors.lastname: Qiu
mesh_terms: D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism; D015153:Blotting, Western; D016203:CDC2 Protein Kinase; D002460:Cell Line; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D015672:Erythroid Precursor Cells / Q000821:virology; D005434:Flow Cytometry; D059565:G2 Phase Cell Cycle Checkpoints / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation; D020411:Oligonucleotide Array Sequence Analysis; D010322:Parvoviridae Infections / Q000378:metabolism*; D016732:Parvovirus B19, Human; D015398:Signal Transduction / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D020687:cdc25 Phosphatases / Q000378:metabolism
chemical_list: C106202:NS1 protein, parvovirus; D017361:Viral Nonstructural Proteins; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases; C517009:CDC25C protein, human; D020687:cdc25 Phosphatases