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Proteome analysis of soluble nuclear proteins reveals that HMGB1/2 suppress genotoxic stress in polyglutamine diseases

Nature Cell Biology volume 9, pages 402–414 (2007)Cite this article

Abstract

Nuclear dysfunction is a key feature of the pathology of polyglutamine (polyQ) diseases. It has been suggested that mutant polyQ proteins impair functions of nuclear factors by interacting with them directly in the nucleus. However, a systematic analysis of quantitative changes in soluble nuclear proteins in neurons expressing mutant polyQ proteins has not been performed. Here, we perform a proteome analysis of soluble nuclear proteins prepared from neurons expressing huntingtin (Htt) or ataxin-1 (AT1) protein, and show that mutant AT1 and Htt similarly reduce the concentration of soluble high mobility group B1/2 (HMGB1/2) proteins. Immunoprecipitation and pulldown assays indicate that HMGBs interact with mutant AT1 and Htt. Immunohistochemistry showed that these proteins were reduced in the nuclear region outside of inclusion bodies in affected neurons. Compensatory expression of HMGBs ameliorated polyQ-induced pathology in primary neurons and in Drosophila polyQ models. Furthermore, HMGBs repressed genotoxic stress signals induced by mutant Htt or transcriptional repression. Thus, HMGBs may be critical regulators of polyQ disease pathology and could be targets for therapy development.

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Figure 1: Proteome analysis of soluble nuclear proteins from primary neurons expressing polyQ implicates HMGB1 and HMGB2 in polyQ pathology.
Figure 2: HMGB1 and HMGB2 proteins interact with mutant polyQ proteins.
Figure 3: HMGB proteins were reduced in neurons of polyQ disease model mice in vivo.
Figure 4: HMGB1 and HMGB2 proteins repress toxicities of mutant Htt and AT1.
Figure 5: HMGB1 represses in vivo toxicity of mutant polyQ proteins in Drosophila models.
Figure 6: Mutant Htt and transcriptional repression similarly induce genotoxic stress signals.
Figure 7: HMGB1 represses genotoxic stress signals induced by mutant Htt and transcriptional repression.
Figure 8: Reduction of HMGB1 induces cell death of primary cortical neurons.

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Acknowledgements

This work was supported by grants to H.O. from the Japan Ministry of Education, Culture, Science, Sports and Technology (16650076), Japan Society for the Promotion of Science (JSPS; 16390249) and Japan Science and Technology Agency (JST; PRESTO). We thank M. Matsumoto and T. Miyashita (Tokyo Metropolitan Institute for Neuroscience, TMIN) for Drosophila experiments, T. Okuda and Shigeki Marubuchi (TMIN, Tokyo Medical and Dental University; TMDU) for primary culture, and T. Tajima for immunohistochemistry. We also thank H. Y. Zoghbi (Baylor College of Medicine) for sharing AT1 knock-in mouse samples and for critical reading of the manuscript.

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Author notes

  1. Kazuhiko Tagawa and Yasushi Enokido: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neuropathology, Medical Research Institute, Tokyo Medical and Dental University, Japan

    Mei-Ling Qi, Kazuhiko Tagawa, Yasushi Enokido, Natsue Yoshimura, Yo-ichi Wada & Hitoshi Okazawa

  2. 21st Century COE Program for Brain Integration and Its Dysfunction, Tokyo Medical and Dental University, Japan

    Mei-Ling Qi, Kei Watase & Hitoshi Okazawa

  3. Faculty of Culture and Science, The University of Tokyo, Japan.,

    Sho-ichi Ishiura

  4. National Center for Neurology and Psychiatry, Japan

    Ichiro Kanazawa

  5. Department of Molecular and Human Genetics, Baylor College of Medicine, USA

    Juan Botas

  6. Department of Molecular Therapeutics, Tokyo Metropolitan Institute for Neuroscience, Japan

    Minoru Saitoe & Hitoshi Okazawa

  7. Department of Neuroproteomics, Max-Delbrück Center for Molecular Medicine, Germany

    Erich E. Wanker

  8. PRESTO, Japan Science and Technology Agency (JST), Japan

    Mei-Ling Qi & Hitoshi Okazawa

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Contributions

This work was mainly performed by M.-L.Q. in collaboration with K.T, Y.E. and other co-authors. The project was planned by H.O.

Corresponding author

Correspondence to Hitoshi Okazawa.

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Supplementary Figures S1, S2, S3, S4, S5, S6, S7 and Supplementary Methods (PDF 1509 kb)

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Qi, ML., Tagawa, K., Enokido, Y. et al. Proteome analysis of soluble nuclear proteins reveals that HMGB1/2 suppress genotoxic stress in polyglutamine diseases. Nat Cell Biol 9, 402–414 (2007). https://doi.org/10.1038/ncb1553

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