title: Benzo[a]pyrene treatment leads to changes in nuclear protein expression and alternative splicing.
abstract: Benzo[a]pyrene (BaP) is a potent pro-carcinogen generated from the combustion of fossil fuel and cigarette smoke. Previously, using a proteomic approach, we have shown that BaP can induce changes in the expression of many cellular proteins, including transcription regulators. In the present study, using a similar approach, we examined the nuclear protein response to BaP in HeLa cells and found that BaP treatment caused expression changes in many nuclear proteins. Twenty-four of these proteins were successfully identified, several of which are involved in the alternative splicing of mRNA, DNA replication, recombination, and repair. The changed expression levels were further confirmed by immunoblot analysis using specific antibodies for two proteins, Lamin A and mitotic checkpoint protein Bub3. The nuclear localization of these two proteins was also confirmed by confocal microscopy. To determine whether alternative splicing was activated following BaP treatment, we examined Fas and CD44, two genes previously shown to be targets of alternative splicing in respond to DNA damage. While no significant activation of alternative splicing was observed for Fas, CD44 splicing variants were found after BaP treatment. Together, these data show that DNA damage induces dramatic changes in nuclear protein expression, and that alternative splicing might be involved in the cellular response to DNA damage.
authors.affiliation: Department of Toxicology, Zhejiang University School of Public Health, 388 Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058, China.
authors.forename: Chunlan
authors.initials: C
authors.lastname: Yan
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Haiyan
authors.initials: H
authors.lastname: Li
authors.forename: Guanglin
authors.initials: G
authors.lastname: Zhang
authors.forename: Penelope J
authors.initials: PJ
authors.lastname: Duerksen-Hughes
authors.forename: Xinqiang
authors.initials: X
authors.lastname: Zhu
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
mesh_terms: D017398:Alternative Splicing / Q000187:drug effects*; D001564:Benzo(a)pyrene / Q000494:pharmacology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002458:Cell Fractionation; D004249:DNA Damage; D008401:Gas Chromatography-Mass Spectrometry; D006367:HeLa Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics; D034904:Lamin Type A / Q000378:metabolism; D009153:Mutagens / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000378:metabolism*; D000075223:Poly-ADP-Ribose Binding Proteins; D019014:fas Receptor / Q000235:genetics
chemical_list: C489444:BUB3 protein, human; D018797:Cell Cycle Proteins; C491901:FAS protein, human; D018960:Hyaluronan Receptors; D034904:Lamin Type A; D009153:Mutagens; D009687:Nuclear Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D019014:fas Receptor; D001564:Benzo(a)pyrene