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Cited by 10DNA Repair MechanismsEffects of Radiation ExposureRadiation Therapy and Dosimetry

A kinome-targeted RNAi-based screen links FGF signaling to H2AX phosphorylation in response to radiation

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Benzina, Sami, Pitaval, Amandine, Lemercier, Claudie, Lustremant, Celine, Frouin, Vincent, Wu, Ning, Papine, Alexandre, Soussaline, Françoise +2 more·2015·Cellular and Molecular Life Sciences
DOI 10.1007/s00018-015-1901-7PMC PMC4548013
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Abstract

A general radioprotective effect by fibroblast growth factor (FGF) has been extensively described since the early 1990s; however, the molecular mechanisms involved remain largely unknown. Radiation-induced DNA double-strand breaks (DSBs) lead to a complex set of responses in eukaryotic cells. One of the earliest consequences is phosphorylation of histone H2AX to form nuclear foci of the phosphorylated form of H2AX (γH2AX) in the chromatin adjacent to sites of DSBs and to initiate the recruitment of DNA-repair molecules. Upon a DSB event, a rapid signaling network is activated to coordinate DNA repair with the induction of cell-cycle checkpoints. To date, three kinases (ATM, ATR, and DNA-PK) have been shown to phosphorylate histone H2AX in response to irradiation. Here, we report a kinome-targeted small interfering RNA (siRNA) screen to characterize human kinases involved in H2AX phosphorylation. By analyzing γH2AX foci at a single-nucleus level, we identified 46 kinases involved either directly or indirectly in H2AX phosphorylation in response to irradiation in human keratinocytes. Furthermore, we demonstrate that in response to irradiation, the FGFR4 signaling cascade promotes JNK1 activation and direct H2AX phosphorylation leading, in turn, to more efficient DNA repair. This can explain, at least partially, the radioprotective effect of FGF.

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  • The selected siRNA dose (20 nM) is used in routine in the authors' lab and is consistent with other works from the literature working with MCF10A or RWPE1.

    “This selected dose is used in routine in our lab (22) and is consistent with other works from the literature working with MCF10A (23) or RWPE1 (24).”

    From Direct transfection of clonal organoids in Matrigel microbeads: a promising approach toward organoid-based genetic screens (2018) · cites this paper as [22]

    The citation (22) is attached to the claim that the citing authors routinely use the selected 20 nM dose; the MCF10A and RWPE1 comparisons have separate citations. The cited paper’s Materials and methods (Standard transfection; siRNA microarray printing; page unavailable) describes siRNA use, including a 20 µM stock solution, but does not report routine use of a 20 nM dose. Searched for the final concentration and routine use. (LLM, conf=0.74)

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Cited by

Sorted by citations

Authors
Benzina, S., Pitaval, A., Lemercier, C., Lustremant, C., Frouin, V., Wu, N., Papine, A., Soussaline, F., Romeo, P.-H., & Gidrol, X.
Affiliations
  • Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
  • Inserm (FR)
  • Imstar (France) (FR)
Corresponding author
Gidrol, Xavier
ORCID
Venue
Cellular and Molecular Life Sciences
Publisher
Springer Science and Business Media LLC
Published
2015-04-18
Volume
72
Issue
18
Pages
3559-3573
ISSN
  • 1420-682X (ISSN-L)
  • 1420-9071
DOI
10.1007/s00018-015-1901-7
PMC
PMC4548013
OpenAlex
W1978082707
Cited by
10
References
2
Citations by year
2023: 1 · 2021: 2 · 2020: 1 · 2018: 3 · 2017: 1 · 2016: 2
FWCI
0.87
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Life Sciences (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL (submittedVersion)
Funders
Agence Nationale de la Recherche (ANR-11-NANB-0002)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Radiation Therapy and Dosimetry
Keywords
FGF signaling, FGFR4, H2AX phosphorylation, γH2AX foci, DNA double-strand breaks, radiation response, DNA repair, kinome screen, radiation protection, keratinocytes, ATM, ATR, DNA-dependent protein kinase, cell cycle checkpoints
MeSH terms
Cell Line, Chromatin, DNA Repair, DNA-Binding Proteins, Fibroblast Growth Factors, Histones, Humans, Nuclear Proteins, Phosphorylation, Radiation, Signal Transduction, Keratinocytes, Cell Cycle Proteins, RNA Interference, RNA, Small Interfering, Mitogen-Activated Protein Kinase 8, Receptor, Fibroblast Growth Factor, Type 4, DNA-Activated Protein Kinase, DNA Breaks, Double-Stranded

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