title: HspA1A facilitates DNA repair in human bronchial epithelial cells exposed to Benzo[a]pyrene and interacts with casein kinase 2.
abstract: Benzo[a]pyrene (BaP) is a ubiquitously distributed environmental pollutant that induces deoxyribonucleic acid (DNA) damage. The inducible heat shock protein (HspA1A) can function as a molecular chaperone; however, its role in DNA repair remains largely unknown. In the present study, human bronchial epithelial cells (16HBE) stably transfected with plasmids carrying HspA1A gene or shRNAs against HspA1A were treated with BaP. DNA damage levels of the cells were evaluated by comet assay. Results suggest that HspA1A could protect cells against DNA damage and facilitate the decrease of DNA damage levels during the first 2 h of DNA repair. DNA repair capacity (DRC) of Benzo(a)pyrene diol epoxide (BPDE)-DNA adducts was evaluated by host cell reactivation assay in the stable 16HBE cells transfected with luciferase reporter vector PCMVluc pretreated with BPDE. Compared with control cells, cells overexpressing HspA1A showed higher DRC (p < 0.01 at 10 μM BPDE and p < 0.05 at 20 μM BPDE, respectively), while knockdown of HspA1A inhibited DNA repair (p < 0.05 at 10 μM BPDE). Moreover, casein kinase 2 (CK2) was shown to interact with HspA1A by mass spectrometry and co-immunoprecipitation assays. The two proteins were co-localized in the cell nucleus and perinuclear region during DNA repair, and were identified by confocal laser scanning microscope. In addition, cells overexpressing HspA1A showed an increased CK2 activity after BaP treatment compared with control cells (p < 0.01). Our results suggest that HspA1A facilitates DNA repair after BaP treatment. HspA1A also interacts with CK2 and enhances the kinase activities of CK2 during DNA repair.
authors: Department of Occupational and Environmental Health and the Ministry of Education Key Lab of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China.
authors: Yanying
authors: Y
authors: Duan
authors: Suli
authors: S
authors: Huang
authors: Jin
authors: J
authors: Yang
authors: Piye
authors: P
authors: Niu
authors: Zhiyong
authors: Z
authors: Gong
authors: Xiaoyong
authors: X
authors: Liu
authors: Lili
authors: L
authors: Xin
authors: R William
authors: RW
authors: Currie
authors: Tangchun
authors: T
authors: Wu
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000378:metabolism; D001564:Benzo(a)pyrene / Q000633:toxicity*; D001980:Bronchi / Q000473:pathology*; D047390:Casein Kinase II / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D019610:Cytoprotection / Q000187:drug effects; D018736:DNA Adducts / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*; D057809:HEK293 Cells; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D014162:Transfection
chemical_list: D018736:DNA Adducts; D018840:HSP70 Heat-Shock Proteins; C490786:HSPA1A protein, human; D001564:Benzo(a)pyrene; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D047390:Casein Kinase II
references: Cell Stress Chaperones. 2006 Autumn;11(3):240-9
references: 17009597
references: Oncogene. 2010 Nov 11;29(45):6016-26
references: 20711232
references: DNA Repair (Amst). 2010 Jul 1;9(7):835-41
references: 20471329
references: Annu Rev Genet. 1988;22:631-77
references: 2853609
references: Radiat Res. 2000 Feb;153(2):186-95
references: 10629618
references: Mol Cells. 2004 Oct 31;18(2):249-55
references: 15529003
references: Cell Stress Chaperones. 2009 Jul;14(4):391-406
references: 19089598
references: Toxicol Lett. 2007 Nov 1;174(1-3):10-7
references: 17900831
references: Trends Cell Biol. 2002 May;12(5):226-30
references: 12062170
references: J Biol Chem. 1998 May 22;273(21):12887-92
references: 9582319
references: Toxicology. 2009 Nov 9;265(1-2):10-4
references: 19748547
references: Science. 1990 May 18;248(4957):850-4
references: 2188360
references: Cell. 1998 Feb 6;92(3):351-66
references: 9476895
references: Cell Stress Chaperones. 2001 Apr;6(2):113-20
references: 11599572
references: J Gen Virol. 1995 Oct;76 ( Pt 10):2423-32
references: 7595346
references: Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2767-71
references: 8146188
references: Cell Stress Chaperones. 2006 Summer;11(2):162-9
references: 16817322
references: Biochim Biophys Acta. 2000 Jul 31;1471(1):M31-41
references: 10967423
references: Cell. 1991 Jul 26;66(2):191-7
references: 1855252
references: J Biol Chem. 2001 Mar 23;276(12):9532-6
references: 11133992
references: J Cell Sci. 2004 Feb 1;117(Pt 4):515-7
references: 14730007
references: Cell Stress Chaperones. 2001 Apr;6(2):136-47
references: 11599575
references: Biomed Environ Sci. 1996 Dec;9(4):370-9
references: 8988805
references: Proteomics. 2005 Aug;5(13):3442-53
references: 16047309
references: Mutat Res. 1993 Aug;294(2):117-26
references: 7687004
references: BMC Mol Biol. 2012 Mar 09;13:7
references: 22404984
references: Cell Stress Chaperones. 2003 Summer;8(2):153-61
references: 14627201
references: Cell Stress Chaperones. 2002 Oct;7(4):396-402
references: 12653484
references: Cell Mol Life Sci. 2009 Jun;66(11-12):1830-9
references: 19387551
references: DNA Repair (Amst). 2009 Jun 4;8(6):682-9
references: 19376751
references: Science. 1989 Oct 20;246(4928):382-5
references: 2799391
references: Exp Cell Res. 1988 Mar;175(1):184-91
references: 3345800
references: Mol Cell. 2012 Feb 10;45(3):371-83
references: 22325354
references: Biochem J. 2003 Jan 1;369(Pt 1):1-15
references: 12396231
references: Cell. 2004 Apr 2;117(1):17-28
references: 15066279
references: Nature. 1988 Apr 28;332(6167):805-10
references: 3282179