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- W2247138151 abstract "C69 DNA-PK, formed by the catalytic subunit DNA-PKcs and the Ku70/Ku86 regulatory heterodimer (Ku) is involved in resistance to cisplatin and Ku reportedly bind cisplatin-DNA adducts. Thus DNA-PK is a molecular target for the improvement of cisplatin therapy and subunits recruited into the DNA-PK protein complex in cisplatin treated cells will be putative molecular targets. In an attempt to identify such putative targets we purified the Ku complex by tandem affinity chromatography before and after cisplatin treatment in HeLa cells and identified changes in its subunits composition through mass spectrometric analysis. We purified the Ku86 complex from HeLa cells untreated or treated for 5 hours with 100I¼g/ml cisplatin and found significant increase of the association of the FACT complex with the Ku86 complex. FACT is composed of two subunits SSRP1 and hspt16 and also binds cisplatin-DNA adducts. We found that endogenous Ku86 and SSRP1 co-immunoprecipitate in MDA-MB-231 cells. Nucleolar subpopulations of Ku86, and SSRP1 exist in the nucleolus of MDA-MB-231 cells. Immunofluorescence analysis shows that Ku86 but not SSRP1 leave the nucleolus after exposure to 1I¼g/ml cisplatin. In contrast, the SSRP1 association with the nucleolus was disrupted by high doses of cisplatin (50-100 I¼g/ml). Thus different regulations may exist controlling the cisplatin-induced losses of nucleolar Ku86 and SSRP1. Furthermore, cisplatin treatment activates DNA-PK and the dissociation of SSRP1 from the nucleolus is suppressed by co-treatment of the cells with Wortmannin or the DNA-PK specific inhibitor NU7026 but not by the PI3K inhibitor LY294002. The cisplatin-induced dissociation of Ku86 from the nucleolus is not suppressed by co-treatment with Kinase inhibitors. These findings suggest a role for DNA-PK in regulating the exit of the FACT complex from the nucleolus after cisplatin treatment and point toward a putative role for FACT in the cellular response to cisplatin." @default.
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- W2247138151 date "2007-11-01" @default.
- W2247138151 modified "2023-09-26" @default.
- W2247138151 title "The DNA-dependent protein kinase (DNA-PK) complex recruits the Facilitator of Chromatin Transcription (FACT) complex and induces the exit of FACT from the nucleolus in response to cisplatin treatment" @default.
- W2247138151 hasPublicationYear "2007" @default.
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