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HspA1A facilitates DNA repair in human bronchial epithelial cells exposed to Benzo[a]pyrene and interacts with casein kinase 2

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Cell Stress and Chaperones Aims and scope

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.

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Acknowledgments

This work was supported by the National Outstanding Youth Science Foundation of China [30525031] and the National Natural Science Foundation of China [30872092].

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Authors and Affiliations

  1. 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

    Yanying Duan, Suli Huang, Jin Yang, Piye Niu, Zhiyong Gong, Xiaoyong Liu, Lili Xin & Tangchun Wu

  2. Department of Occupational and Environmental Health, School of Public Health, Xiangya Medical College, Central South University, Changsha, 410078, Hunan, China

    Yanying Duan

  3. Department of Anatomy and Neurobiology, Dalhousie University, 5850 College Street, Halifax, Nova Scotia, B3H 4R2, Canada

    R. William Currie

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  1. Yanying Duan
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  2. Suli Huang
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  3. Jin Yang
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  9. Tangchun Wu
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Correspondence to Tangchun Wu.

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Yanying Duan and Suli Huang contributed equally to this work

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Duan, Y., Huang, S., Yang, J. et al. HspA1A facilitates DNA repair in human bronchial epithelial cells exposed to Benzo[a]pyrene and interacts with casein kinase 2. Cell Stress and Chaperones 19, 271–279 (2014). https://doi.org/10.1007/s12192-013-0454-7

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  • DOI: https://doi.org/10.1007/s12192-013-0454-7

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