Dataset Information

MS-based proteomic identification of NLRP3 binding partners in HeLa cells

ABSTRACT: DNA double strand break (DSB) is a highly toxic lesion that can generate genome instability, a major source of tumorigenesis. DSBs are mainly repaired by nonhomologous end joining (NHEJ) or homologous recombination (HR). The selection of the DSB repair pathway primarily depends on the DNA resection of the DSB ends. Indeed, HR is initiated by resection at the DSB generating 3' single stranded extension. The shieldin complex prevents resection fostering DSB repair toward NHEJ. In the context of genome stability, NLRP3 was recently reported to interact with ATM and to enhance its activation in response to DNA DSBs. Whether NLRP3 controls the DSB repair machinery remains unknown. To further elucidate the function of NLRP3 in DNA repair, we sought NLRP3-binding partners by screening proteins that bind to FLAG-tagged NLRP3 (FLAG-NLRP3) in HeLa cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Yohann Couté

LAB HEAD: Yohann Couté

PROVIDER: PXD058288 | Pride | 2025-07-14

REPOSITORIES: Pride

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Publications

The inflammasome sensor NLRP3 interacts with REV7 to maintain genome integrity through homologous recombination.

Burlet Delphine D Khan Md Muntaz MM Hacot Sabine S Buthmann Hannes H Bardoulet Léa L Huber Anne-Laure AL Gorry Julie J Föhr Bastian B Lopez Bernard S BS Couté Yohann Y Faesen Alex C AC Geyer Matthias M Tissier Agnès A Petrilli Virginie V

Nucleic acids research 20250601 12

DNA double-strand break (DSB) is a highly toxic lesion that can generate genome instability, a major source of tumorigenesis. DSBs are mainly repaired by non-homologous end joining (NHEJ) or homologous recombination (HR). The selection of the DSB repair pathway primarily depends on the DNA resection of the DSB ends. Indeed, HR is initiated by resection at the DSB, generating 3' single-stranded overhang. The shieldin complex prevents resection fostering DSB repair toward NHEJ. Here, we reveal tha ...[more]

PMID: 40613708

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Project description:The proposal aims to characterise the pathways of DSB repair and recombination in Arabidopsis with the main focus of this research being the NHEJ pathway of illegitimate recombination. We will build on our expertise and resources in the field of DSB repair in plants, using the Arabidopsis NHEJ mutant atku80 which is an excellent model system for the study of DSB repair in higher eukaryotes (West et al., 2002 Plant J. 31, 517-28). Comparison of the transcriptome in NHEJ mutant and wild type plants under different experimental conditions will identify novel candidate genes involved in DNA DSB repair or damage signalling pathways.We will conduct 4 separate experiments on Arabidopsis seedlings grown on 0.5 MS media: the first will be a control consisting of Wassilewskija (WS-2) plants grown under standard conditions. In the second experiment WS plants will be exposed to the chemotherapeutic agent bleomycin (1 microgram per ml). This agent has been shown to cause both single and double strand breaks in the genome of Arabidopsis seedlings (Menke et al., 2001 Mut. Res. 493, 87-93). This agent is used in preference to gamma irradiation, which causes high levels of oxidative damage to cellular components. These experiments will characterise for the first time the transcriptional responses of Arabidopsis cells to the specific induction of DNA strand breaks. The transcript profile will also be determined in untreated atku80 mutants. This experiment will identify the components of novel and previously characterised DNA repair pathways up regulated in the mutant background. Finally, transcript analysis of bleomycin treated atku80 mutants and comparison with untreated mutant plants and both untreated and treated WS plants will identify novel putative DSB repair genes up regulated in response to genotoxic stress in the absence of a Ku80 mediated DSB repair pathway.The results of these studies will provide new and important information which will be instrumental in identifying novel genes and pathways involved in DNA repair and genome stability in plants and help elucidate the fundamental differences that exist between animal and plant DSB repair processes. Experiment Overall Design: Number of plants pooled:20

			Action	DRS
	FLAG-NLRP3_R1.mgf	Mgf
	FLAG-NLRP3_R1.raw	Raw
	FLAG-NLRP3_R2.mgf	Mgf
	FLAG-NLRP3_R2.raw	Raw
	FLAG-NLRP3_R3.mgf	Mgf