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Ars2 maintains neural stem-cell identity through direct transcriptional activation of Sox2

Abstract

Fundamental questions remain unanswered about the transcriptional networks that control the identity and self-renewal of neural stem cells (NSCs), a specialized subset of astroglial cells that are endowed with stem properties and neurogenic capacity. Here we report that the zinc finger protein Ars2 (arsenite-resistance protein 2; also known as Srrt) is expressed by adult NSCs from the subventricular zone (SVZ) of mice, and that selective knockdown of Ars2 in cells expressing glial fibrillary acidic protein within the adult SVZ depletes the number of NSCs and their neurogenic capacity. These phenotypes are recapitulated in the postnatal SVZ of hGFAP-cre::Ars2fl/fl conditional knockout mice, but are more severe. Ex vivo assays show that Ars2 is necessary and sufficient to promote NSC self-renewal, and that it does so by positively regulating the expression of Sox2. Although plant1,2,3 and animal4,5 orthologues of Ars2 are known for their conserved roles in microRNA biogenesis, we unexpectedly observed that Ars2 retains its capacity to promote self-renewal in Drosha and Dicer1 knockout NSCs. Instead, chromatin immunoprecipitation revealed that Ars2 binds a specific region within the 6-kilobase NSC enhancer of Sox2. This association is RNA-independent, and the region that is bound is required for Ars2-mediated activation of Sox2. We used gel-shift analysis to refine the Sox2 region bound by Ars2 to a specific conserved DNA sequence. The importance of Sox2 as a critical downstream effector is shown by its ability to restore the self-renewal and multipotency defects of Ars2 knockout NSCs. Our findings reveal Ars2 as a new transcription factor that controls the multipotent progenitor state of NSCs through direct activation of the pluripotency factor Sox2.

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Figure 1: Ars2 maintains neural stem cells in the adult SVZ.
Figure 2: Ars2 regulates postnatal neurogenesis.
Figure 3: Ars2 acts independently of the miRNA pathway to promote NSC self-renewal through Sox2.
Figure 4: Ars2 directly activates transcription of Sox2 to mediate NSC self-renewal and multipotency.

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Acknowledgements

We thank X. Lu, M. Götz, A. Rizzino, P. M. LLedo, P. Charneau, M. Segura, P. L. Howard and S. Olejniczak for reagents. We are grateful to K. Hadjantonakis, A. Ferrer-Vaquer, J. Zhang and Y. Ganat for assistance. U. Ruthishauser, V. Tabar and the Molecular Cytology Core Facility at the Memorial Sloan-Kettering Cancer Center shared equipment. S. R. Ferron, H. Mira, A. Joyner, H. Duan, Q. Dai, I. Farinas and S. Shi provided critical comments. Work in E.C.L.’s group was supported by the Burroughs Wellcome Fund, the Starr Cancer Consortium (I3-A139) and the NIH (R01-GM083300). C.A.-A. was funded by a EMBO Long-Term Fellowship (ALTF 718-2008) and a NYSTEM Fellowship.

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C.A.-A. performed and designed all of the experiments, T.M. performed in vivo lentivirus injections and C.B.T. provided reagents. C.A.-A. and E.C.L. conceived the project, interpreted the results and wrote the manuscript.

Corresponding authors

Correspondence to Celia Andreu-Agullo or Eric C. Lai.

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The authors declare no competing financial interests.

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Andreu-Agullo, C., Maurin, T., Thompson, C. et al. Ars2 maintains neural stem-cell identity through direct transcriptional activation of Sox2. Nature 481, 195–198 (2012). https://doi.org/10.1038/nature10712

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