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.

Equine Epidermis: A Source of Epithelial-Like Stem/Progenitor Cells with In Vitro and In Vivo Regenerative Capacities

“…Color images available online at www.liebertpub.com/scd expression in corneal stem cells [41,42] and circulating keratinocyte progenitor cells [32]. Moreover, the EpSCs in the present study were positive for Casein Kinase 2b, which is reputed to inhibit the development of the mesenchymal morphology and phenotype [37,45], and plays an important role in the functional regulation of epithelial cells [46][47][48].…”

DEMETER plant DNA demethylase induces antiviral response by interferon signalling in animal cells

“…Many Hsps are upregulated in response to a variety of stress stimuli including DNA damage 26 , 27 , and we observed a substantial increase in the expression levels of diverse Hsp genes in the 293T-DMEΔ cells (Table 1 and Supplementary Fig. 4b ).…”

TRiC controls transcription resumption after UV damage by regulating Cockayne syndrome protein A

“…By regulating protein stability and degradation in cells, the PN drives vital processes 30 . Although several components of the PN have been found implicated in the DNA damage response 31 – 34 , mechanistic insight into how this network affects these processes has remained largely elusive. Here we demonstrate that one of the components of the PN, the chaperonin TRiC, stably interacts with the core TC-NER protein CSA.…”