title: Engagement of Components of DNA-Break Repair Complex and NFκB in Hsp70A1A Transcription Upregulation by Heat Shock.
abstract: An involvement of components of DNA-break repair (DBR) complex including DNA-dependent protein kinase (DNA-PK) and poly-ADP-ribose polymerase 1 (PARP-1) in transcription regulation in response to distinct cellular signalling has been revealed by different laboratories. Here, we explored the involvement of DNA-PK and PARP-1 in the heat shock induced transcription of Hsp70A1A. We find that inhibition of both the catalytic subunit of DNA-PK (DNA-PKc), and Ku70, a regulatory subunit of DNA-PK holo-enzyme compromises transcription of Hsp70A1A under heat shock treatment. In immunoprecipitation based experiments we find that Ku70 or DNA-PK holoenzyme associates with NFκB. This NFκB associated complex also carries PARP-1. Downregulation of both NFκB and PARP-1 compromises Hsp70A1A transcription induced by heat shock treatment. Alteration of three bases by site directed mutagenesis within the consensus κB sequence motif identified on the promoter affected inducibility of Hsp70A1A transcription by heat shock treatment. These results suggest that NFκB engaged with the κB motif on the promoter cooperates in Hsp70A1A activation under heat shock in human cells as part of a DBR complex including DNA-PK and PARP-1.
authors.affiliation: Division of Molecular Medicine, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata, India.
authors.forename: Joyita
authors.initials: J
authors.lastname: Hazra
authors.affiliation: Division of Molecular Medicine, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata, India.
authors.forename: Pooja
authors.initials: P
authors.lastname: Mukherjee
authors.affiliation: Division of Molecular Medicine, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata, India.
authors.forename: Asif
authors.initials: A
authors.lastname: Ali
authors.affiliation: Bioinformatics Center, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata, India.
authors.forename: Soumita
authors.initials: S
authors.lastname: Poddar
authors.affiliation: Division of Molecular Medicine, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata, India.
authors.forename: Mahadeb
authors.initials: M
authors.lastname: Pal
mesh_terms: D020134:Catalytic Domain / Q000235:genetics; D045744:Cell Line, Tumor; D053960:DNA Breaks; D004260:DNA Repair / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D051747:DNA-Activated Protein Kinase / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D000072200:Ku Autoantigen / Q000037:antagonists & inhibitors; D052002:NF-kappa B p50 Subunit / Q000235:genetics*; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051996:Transcription Factor RelA / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D018840:HSP70 Heat-Shock Proteins; D052002:NF-kappa B p50 Subunit; C498143:RELA protein, human; D034741:RNA, Small Interfering; D051996:Transcription Factor RelA; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D051747:DNA-Activated Protein Kinase; D000072200:Ku Autoantigen; D004250:DNA Topoisomerases, Type II