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Neurogenesis requires TopBP1 to prevent catastrophic replicative DNA damage in early progenitors

Abstract

The rapid proliferation of progenitors during neurogenesis requires a stringent genomic maintenance program to ensure transmission of genetic fidelity. However the essential factors that govern neural progenitor genome integrity are unknown. Here we report that conditional inactivation of mouse TopBP1, a protein linked to DNA replication, and a key activator of the DNA damage response kinase ATR (ataxia telangiectasia and rad3-related) is critical for maintenance of early-born neural progenitors. During cortical development TopBP1 prevented replication-associated DNA damage in Emx1-progenitors which otherwise resulted in profound tissue ablation. Notably, disrupted neurogenesis in TopBP1-depleted tissues was substantially rescued by inactivation of p53 but not of ATM. Our data establish that TopBP1 is essential for preventing replication-associated DNA strand breaks, but is not essential per se for DNA replication. Thus, TopBP1 is crucial for maintaining genome integrity in the early progenitors that drive neurogenesis.

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Figure 1: Topbp1 deletion in the nervous system.
Figure 2: Apoptosis and cortical layering disruption in the Topbp1Nes-cre brain.
Figure 3: Dorsal telencephalon progenitors are lost in Topbp1Emx1-cre mice.
Figure 4: Analysis of cortical development in Lig4Emx1-cre and Xrcc1Emx1-cre brain.
Figure 5: Defective neurogenesis after Topbp1 inactivation requires p53 but not ATM signaling.
Figure 6: TopBP1 deficiency during neurogenesis results in DNA strand break accumulation in cortical progenitors.
Figure 7: Analysis of DNA damage responses in TopBP1-depleted cells.

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Acknowledgements

We thank the Hartwell Center for biotech support, the Transgenic Core Facility for blastocyst injections and chimera production and the ARC for animal husbandry. P.J.M. is supported by the US National Institutes of Health (NS-37956, CA-21765), a Cancer Center support grant (P30 CA21765) and the American Lebanese and Syrian Associated Charities of St. Jude Children's Research Hospital. S.K. is a Neoma Boadway Academic Programs Endowed Fellow.

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Y.L., S.K. and H.R.R. performed all experiments characterizing the Topbp1 mutant mouse and contributed to writing the manuscript. S.M.D. contributed the analysis of the Lig4 and Xrcc1 conditional mutant mice. J.Z. generated western blot data. P.J.M. was project leader and produced the final version of the manuscript.

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Correspondence to Peter J McKinnon.

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Lee, Y., Katyal, S., Downing, S. et al. Neurogenesis requires TopBP1 to prevent catastrophic replicative DNA damage in early progenitors. Nat Neurosci 15, 819–826 (2012). https://doi.org/10.1038/nn.3097

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