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Research
Dynamic Alterations of Histone H3 Phospho-acetylation Correlate With Radio Sensitivity of Mitotic Cells During DNA Damage
Sanjay Gupta
Epigenetics and Chromatin Biology Group, Gupta Lab, Cancer Research Institute, Advanced Centre for Treatment, Research and Education in Cancer, Tata Memorial Centre, Kharghar, Navi Mumbai- 410210,MH, India
Corresponding Author
ORCiD: https://orcid.org/0000-0002-1209-189X
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This work is licensed under a CC BY 4.0 License. Read Full License
Background - Histone Post Translational Modifications (PTMs) change in a cell cycle dependent manner and also orchestrate the DNA repair process for radiation induced DNA damage. Mitosis is the most radiosensitive phase of the cell cycle but the epigenetic events that regulate its radiosensitivity remain elusive.
Results - This study explored the dynamics between histone marks H3S10/S28ph, H3K9ac and γH2AX during mitotic DNA damage response. The presence of a mononucleosome level association between γH2AX and H3S10ph was observed only during mitosis. This association was abrogated upon cell cycle progression and chromatin de-condensation, concomitant with chromatin recruitment of DNA repair proteins Ku70 and Rad51. Moreover, the levels of H3S10/28ph remained unchanged upon DNA damage during mitosis, but decreased in a cell cycle dependent manner upon mitotic exit. However, the population that arose after mitotic progression of damaged cells comprised of binucleated tetraploid cells. This population was epigenetically distinct from interphase cells, characterized by reduced H3S10/S28ph, increased H3K9ac and more open chromatin configuration. These epigenetic features correlated with decreased survival potential of this population. The low levels of H3S10/28ph were attributed to decreased protein translation and chromatin recruitment of histone kinase Mitogen and Stress-activated Kinase 1 (MSK1) along with persistent levels of Protein phosphatase1 catalytic subunit α (PP1α).
Conclusions – This study suggests that a unique epigenetic landscape attained during and after mitotic DNA damage collectively contributed to mitotic radiosensitivity. The findings of this study have potential clinical significance in terms of tackling resistance against anti-mitotic chemotherapeutic agents.
Keywords
Mitosis, Histone Post Translational Modifications, DNA damage, Radiosensitivity
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- AdditionalFile3.tiff
Additional File 3 Controls for Mononucleosomal Immunoprecipitation (A) TAE-Agarose gel based representation of mononucleosomes prepared from IR exposed mitosis synchronized MCF7 cells. 0 and 4 indicate time (hours) chosen after radiation and nocodazole release for carrying out immunoprec ipitation experiment. (B) Validation of anti-γH2AX and IgG anti-mouse antibody and (C) anti-H3S10ph and IgG anti-rabbit antibody by immuno-precipitation and immuno-blotting. L- 100bp ladder, b.p. – base pair,IP-Immuno-precipitation,IB-Immunoblotting
- AdditionalFile2.tiff
Additional File 2 Effect of different DNA damaging agents, cell lines and radiation doses in response to DNA damage in mitosis. (A) Western blotting for histone PTMs γH2AX, H3S10ph, H3S28ph and H3K9ac from acid extracted histones of mitosis synchronized MCF7 cells treated with cisplatin, H2O2, Adriamycin and UV rays, (B) different doses of ionizing radiation to induce DNA damage. (C) U87 glioblastoma and (D) AGS gastric adenocarcinoma cell line. Histone H3 serves as loading control. Hrs. – Hours, Gy- Gray.
- AdditionalFile1.tiff
Additional File 1 Histone PTM alterations during mitotic DDR Representative z-stack projection images for co-immunofluorescence of γH2AX with histone PTMs (A) H3S10ph, (B) H3S28ph and (C) H3K9ac at time points 0, 4 and 24 hours after radiation and nocodazole release. Scale bar for each image is 10μm. DAPI staining depicts nucleus. Hrs.- Hours.
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Research
Dynamic Alterations of Histone H3 Phospho-acetylation Correlate With Radio Sensitivity of Mitotic Cells During DNA Damage
Sanjay Gupta
Epigenetics and Chromatin Biology Group, Gupta Lab, Cancer Research Institute, Advanced Centre for Treatment, Research and Education in Cancer, Tata Memorial Centre, Kharghar, Navi Mumbai- 410210,MH, India
Corresponding Author
ORCiD: https://orcid.org/0000-0002-1209-189X
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 23 Sep, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Epigenetics & Genomics
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background - Histone Post Translational Modifications (PTMs) change in a cell cycle dependent manner and also orchestrate the DNA repair process for radiation induced DNA damage. Mitosis is the most radiosensitive phase of the cell cycle but the epigenetic events that regulate its radiosensitivity remain elusive.
Results - This study explored the dynamics between histone marks H3S10/S28ph, H3K9ac and γH2AX during mitotic DNA damage response. The presence of a mononucleosome level association between γH2AX and H3S10ph was observed only during mitosis. This association was abrogated upon cell cycle progression and chromatin de-condensation, concomitant with chromatin recruitment of DNA repair proteins Ku70 and Rad51. Moreover, the levels of H3S10/28ph remained unchanged upon DNA damage during mitosis, but decreased in a cell cycle dependent manner upon mitotic exit. However, the population that arose after mitotic progression of damaged cells comprised of binucleated tetraploid cells. This population was epigenetically distinct from interphase cells, characterized by reduced H3S10/S28ph, increased H3K9ac and more open chromatin configuration. These epigenetic features correlated with decreased survival potential of this population. The low levels of H3S10/28ph were attributed to decreased protein translation and chromatin recruitment of histone kinase Mitogen and Stress-activated Kinase 1 (MSK1) along with persistent levels of Protein phosphatase1 catalytic subunit α (PP1α).
Conclusions – This study suggests that a unique epigenetic landscape attained during and after mitotic DNA damage collectively contributed to mitotic radiosensitivity. The findings of this study have potential clinical significance in terms of tackling resistance against anti-mitotic chemotherapeutic agents.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8