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Deubiquitylase HAUSP stabilizes REST and promotes maintenance of neural progenitor cells

Nature Cell Biology volume 13, pages 142–152 (2011)Cite this article

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Abstract

The repressor element 1-silencing transcription factor (REST) functions as a master regulator to maintain neural stem/progenitor cells (NPCs). REST undergoes proteasomal degradation through β-TrCP-mediated ubiquitylation during neuronal differentiation. However, reciprocal mechanisms that stabilize REST in NPCs are undefined. Here we show that the deubiquitylase HAUSP counterbalances REST ubiquitylation and prevents NPC differentiation. HAUSP expression declines concordantly with REST on neuronal differentiation and reciprocally with β-TrCP levels. HAUSP knockdown in NPCs decreases REST and induces differentiation. In contrast, HAUSP overexpression upregulates REST by overriding β-TrCP-mediated ubiquitylation. A consensus site (310-PYSS-313) in human REST is required for HAUSP-mediated REST deubiquitylation. Furthermore, REST overexpression in NPCs rescues the differentiation phenotype induced by HAUSP knockdown. These data demonstrate that HAUSP stabilizes REST through deubiquitylation and antagonizes β-TrCP in regulating REST at the post-translational level. Thus, HAUSP-mediated deubiquitylation represents a critical regulatory mechanism involved in the maintenance of NPCs.

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Figure 1: HAUSP and REST protein levels decline coordinately during neuronal differentiation.
Figure 2: HAUSP knockdown reduces REST protein levels in NPCs.
Figure 3: HAUSP knockdown promotes neural differentiation and decreases NPC self-renewal, and REST overexpression rescues the differentiation phenotype induced by HAUSP knockdown.
Figure 4: rtPCR analysis indicated that reduced HAUSP expression by shRNA did not significantly alter REST mRNA expression, but increased TUJ1 (a REST target gene) mRNA levels.
Figure 5: HAUSP mediates REST deubiquitylation to regulate REST protein levels.
Figure 6: Neuronal differentiation induced by knockdown of endogenous HAUSP was rescued by ectopic expression of wild-type HAUSP, but not the catalytic dead HAUSP mutant.
Figure 7: A consensus site of human REST (310-PYSS-313) is required for the HAUSP-mediated REST deubiquitylation and HAUSP counteracts β-TrCP-mediated REST ubiquitylation.
Figure 8: ‘Ying–Yang’ control model of REST protein at the post-translational level.

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Acknowledgements

We thank members in the J.N.R. lab for helpful discussions and suggestions. We are also grateful to the Flow Cytometry Core, Imaging Core and Central Cell Services at Cleveland Clinic Lerner Research Institute for their help and services. This work was supported by a research fund from Cleveland Clinic Foundation and a NIH grant (NS070315) to S.B.

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Authors and Affiliations

  1. Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, NE30, Cleveland, Ohio 44195, USA

    Zhi Huang, Qiulian Wu, Olga A. Guryanova, Lin Cheng, Jeremy N. Rich & Shideng Bao

  2. Center for Experimental Research, The First People’s Hospital, Shanghai Jiaotong University, Shanghai 200080, China

    Lin Cheng

  3. Department of Pediatrics, Riley Heart Research Center, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA

    Weinian Shou

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Contributions

Z.H., Q.W., O.A.G. and L.C. carried out and planned all experiments. S.B. developed the hypothesis, coordinated the study, oversaw the research and results, and wrote the manuscript. J.N.R. helped to write the manuscript and provided input into design and interpretation. W.S. provided reagents and helpful suggestions. The work was carried out in the laboratory of S.B.

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Correspondence to Shideng Bao.

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Huang, Z., Wu, Q., Guryanova, O. et al. Deubiquitylase HAUSP stabilizes REST and promotes maintenance of neural progenitor cells. Nat Cell Biol 13, 142–152 (2011). https://doi.org/10.1038/ncb2153

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