Abstract
The INO80 (inositol requiring mutant 80) chromatin remodeling complex plays important roles in transcriptional regulation and DNA replication and repair, and consists of several functional protein subunits, including the critical Ino80 ATPase catalytic subunit. While the function of INO80 has been studied in yeast and mammalian cell lines, we do not know how mIno80 contributes to the maintenance of genome stability to prevent cancer development in mice. Here, we use a conditional knockout approach to explore the cellular and organismal functions of mIno80. Deletion of mIno80 results in profound cellular proliferative defects and activation of p21-dependent cellular senescence. While mIno80 is required for efficient repair of DNA double strand breaks, its depletion did not impact upon the formation of γ-H2AX and 53BP1 DNA damage foci, or the activation of the ATM-CHK2-dependent DNA damage response. mIno80 deletion inhibited the generation of single-strand DNA, resulting in defects in homology-directed DNA repair (HDR) at telomeres. Fragile telomeres were prominent in mIno80Δ/Δ MEFs, suggesting that chromatin remodeling is required for efficient telomere replication. mIno80−/− mouse embryos die early during embryogenesis, while conditional deletion of mIno80 in adult mice results in weight loss and premature death. In a p53−/− tumor-prone background, mIno80 haploinsufficiency favored the development of sarcomas. Our studies suggest that the mIno80 chromatin remodeling complex plays important roles in telomere replication, HDR-mediated repair of dysfunctional telomeres, and maintenance of genome stability.
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Acknowledgements
This work was supported by funding from NCI (RO1 CA129037 to SC; CA127945 and CA097175 Project 3 to LL).
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Supplementary information, Figure S1
Deletion of mouse mIno80 gene. (PDF 452 kb)
Supplementary information, Figure S2
DNA damage foci are present in mIno80 deleted cells. (PDF 704 kb)
Supplementary information, Figure S3
(A) Representative colony forming assays of 4-HT treated CAG-CreER; mIno80+/+ and CAG-CreER; mIno80F/F MEFs following exposure to 0.2μM aphidicolin (APH) or 0.05mM hydroxyurea for 24hrs. (PDF 3242 kb)
Supplementary information, Figure S4
mIno80 deletion did not alter total telomere length, G-overhang status, and recruitment 53BP1 to dysfunctional telomeres. (PDF 1349 kb)
Supplementary information, Figure S5
mIno80 deletion does not impact upon C-NHEJ or A-NHEJ mediated DNA repair. (PDF 1548 kb)
Supplementary information, Figure S6
Phenotype of mIno80 heterozygote mice. (PDF 3742 kb)
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Min, JN., Tian, Y., Xiao, Y. et al. The mINO80 chromatin remodeling complex is required for efficient telomere replication and maintenance of genome stability. Cell Res 23, 1396–1413 (2013). https://doi.org/10.1038/cr.2013.113
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DOI: https://doi.org/10.1038/cr.2013.113
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