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Parvovirus B19 NS1 protein induces cell cycle arrest at G2-phase by activating the ATR-CDC25C-CDK1 pathway.

PLoS Pathog.. 2017; 
XuPeng,ZhouZhe,XiongMin,ZouWei,DengXuefeng,GanaieSafder S,KleiboekerSteve,PengJianxin,LiuKaiyu,WangShengqi,YeShui Qing,QiuJian
Products/Services Used Details Operation
The following first antibodies were purchased: anti-Strep (A01732) from GenScript, Piscataway, NJ; mouse anti-BrdU (clone B44) (347580), anti-cyclin B1 (554179), anti-CDK1 (pY15) (612306) from BD, San Jose, CA; rabbit anti-BrdU (600-401-C29) and mouse anti-Flag (2001301-1313) from Rockland, Limerick, PA; anti-CDK1 (A303-663A), anti-CDC25C (A301390A), anti-MYT1 (A302-424A), anti-p38 (A300-707A) from Bethyl, Montgomery, TX; antiCDK1 (pT161) (P12-1090) from Assay Biotech, Sunnyvale, CA; anti-CDC25C(pS216) (4204), anti-γH2AX(pS139) (05–636), anti-B19V capsid (MAB8292) from Millipore, Billerica, MA; anti-WEE1 (AP8106b), anti-CHK1 (AM7401a), anti-CHK2 (AP4999a), anti-MAPKAPK2 (pS272) (AP3147a) from Abgent, San Diego, CA; anti-ATR (ab2905), anti-GAPDH (Ab8245) from Abcam, Cambridge, MA; anti-ATR (pT1989) (GTX128145), anti-CHK1(pS345) (GTX100065S), anti-RPA32(pT21) (GTX62664) from GeneTex, Irvine, CA; anti-ATM (2873), anti-MAPKAPK2(MK2) (12155s), anti-MARK3 (9311s), anti-p21 (2947p), anti-Histone H3 (9715S) from Cell Signaling Technology, Danvers, MA; anti-ATM(pS1981) (2952–1), antiCHK2(pS33/35) (2297–1) from Epitomics, Burlingame, CA; anti-p38 (pT180/Y182) (p1901802) from Phosphosolutions, Aurora, CO; anti-β-actin (A5441) from Sigma-Aldrich, St. Louis, MO.

摘要

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 T... More

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