Mutant huntingtin binds the mitochondrial fission GTPase dynamin-related protein-1 and increases its enzymatic activity (original) (raw)

Nature Medicine volume 17, pages 377–382 (2011)Cite this article

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Abstract

Huntington's disease is an inherited and incurable neurodegenerative disorder caused by an abnormal polyglutamine (polyQ) expansion in huntingtin (encoded by HTT). PolyQ length determines disease onset and severity, with a longer expansion causing earlier onset. The mechanisms of mutant huntingtin-mediated neurotoxicity remain unclear; however, mitochondrial dysfunction is a key event in Huntington's disease pathogenesis1,2. Here we tested whether mutant huntingtin impairs the mitochondrial fission-fusion balance and thereby causes neuronal injury. We show that mutant huntingtin triggers mitochondrial fragmentation in rat neurons and fibroblasts of individuals with Huntington's disease in vitro and in a mouse model of Huntington's disease in vivo before the presence of neurological deficits and huntingtin aggregates. Mutant huntingtin abnormally interacts with the mitochondrial fission GTPase dynamin-related protein-1 (DRP1) in mice and humans with Huntington's disease, which, in turn, stimulates its enzymatic activity. Mutant huntingtin–mediated mitochondrial fragmentation, defects in anterograde and retrograde mitochondrial transport and neuronal cell death are all rescued by reducing DRP1 GTPase activity with the dominant-negative DRP1 K38A mutant. Thus, DRP1 might represent a new therapeutic target to combat neurodegeneration in Huntington's disease.

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References

  1. Bossy-Wetzel, E., Petrilli, A. & Knott, A.B. Mutant huntingtin and mitochondrial dysfunction. Trends Neurosci. 31, 609–616 (2008).
    Article CAS Google Scholar
  2. Lin, M.T. & Beal, M.F. Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443, 787–795 (2006).
    Article CAS Google Scholar
  3. Knott, A.B. & Bossy-Wetzel, E. Impairing the mitochondrial fission and fusion balance: a new mechanism of neurodegeneration. Ann. NY Acad. Sci. 1147, 283–292 (2008).
    Article CAS Google Scholar
  4. Knott, A.B., Perkins, G., Schwarzenbacher, R. & Bossy-Wetzel, E. Mitochondrial fragmentation in neurodegeneration. Nat. Rev. Neurosci. 9, 505–518 (2008).
    Article CAS Google Scholar
  5. Chen, H. et al. Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J. Cell Biol. 160, 189–200 (2003).
    Article CAS Google Scholar
  6. Smirnova, E., Griparic, L., Shurland, D.L. & van der Bliek, A.M. Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol. Biol. Cell 12, 2245–2256 (2001).
    Article CAS Google Scholar
  7. Wang, H., Lim, P.J., Karbowski, M. & Monteiro, M.J. Effects of overexpression of huntingtin proteins on mitochondrial integrity. Hum. Mol. Genet. 18, 737–752 (2009).
    Article CAS Google Scholar
  8. Barsoum, M.J. et al. Nitric oxide–induced mitochondrial fission is regulated by dynamin-related GTPases in neurons. EMBO J. 25, 3900–3911 (2006).
    Article CAS Google Scholar
  9. Liot, G. et al. Complex II inhibition by 3-NP causes mitochondrial fragmentation and neuronal cell death via an NMDA- and ROS-dependent pathway. Cell Death Differ. 16, 899–909 (2009).
    Article CAS Google Scholar
  10. Yuan, H. et al. Mitochondrial fission is an upstream and required event for bax foci formation in response to nitric oxide in cortical neurons. Cell Death Differ. 14, 462–471 (2007).
    Article CAS Google Scholar
  11. Gauthier, L.R. et al. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules. Cell 118, 127–138 (2004).
    Article CAS Google Scholar
  12. Orr, A.L. et al. N-terminal mutant huntingtin associates with mitochondria and impairs mitochondrial trafficking. J. Neurosci. 28, 2783–2792 (2008).
    Article CAS Google Scholar
  13. Hodgson, J.G. et al. Human huntingtin derived from YAC transgenes compensates for loss of murine huntingtin by rescue of the embryonic lethal phenotype. Hum. Mol. Genet. 5, 1875–1885 (1996).
    Article CAS Google Scholar
  14. Slow, E.J. et al. Selective striatal neuronal loss in a YAC128 mouse model of Huntington disease. Hum. Mol. Genet. 12, 1555–1567 (2003).
    Article CAS Google Scholar
  15. Perkins, G.A. et al. Electron tomography of mitochondria after the arrest of protein import associated with Tom19 depletion. Eur. J. Cell Biol. 80, 139–150 (2001).
    Article CAS Google Scholar
  16. Cui, L. et al. Transcriptional repression of PGC-1α by mutant huntingtin leads to mitochondrial dysfunction and neurodegeneration. Cell 127, 59–69 (2006).
    Article CAS Google Scholar
  17. Fan, J., Cowan, C.M., Zhang, L.Y., Hayden, M.R. & Raymond, L.A. Interaction of postsynaptic density protein-95 with NMDA receptors influences excitotoxicity in the yeast artificial chromosome mouse model of Huntington′s disease. J. Neurosci. 29, 10928–10938 (2009).
    Article CAS Google Scholar
  18. Graham, R.K. et al. Differential susceptibility to excitotoxic stress in YAC128 mouse models of Huntington disease between initiation and progression of disease. J. Neurosci. 29, 2193–2204 (2009).
    Article CAS Google Scholar
  19. Cho, D.H. et al. _S_-nitrosylation of Drp1 mediates β-amyloid–related mitochondrial fission and neuronal injury. Science 324, 102–105 (2009).
    Article CAS Google Scholar
  20. Bossy, B. et al. _S_-nitrosylation of DRP1 does not affect enzymatic activity and is not specific to Alzheimer′s disease. J. Alzheimers Dis. 20 (Suppl 2), S513–S526 (2010).
    Article Google Scholar
  21. Kaltenbach, L.S. et al. Huntingtin interacting proteins are genetic modifiers of neurodegeneration. PLoS Genet. 3, e82 (2007).
    Article Google Scholar
  22. Li, S.H. & Li, X.J. Huntingtin-protein interactions and the pathogenesis of Huntington′s disease. Trends Genet. 20, 146–154 (2004).
    Article Google Scholar
  23. Choo, Y.S., Johnson, G.V., MacDonald, M., Detloff, P.J. & Lesort, M. Mutant huntingtin directly increases susceptibility of mitochondria to the calcium-induced permeability transition and cytochrome c release. Hum. Mol. Genet. 13, 1407–1420 (2004).
    Article CAS Google Scholar
  24. Panov, A.V. et al. Early mitochondrial calcium defects in Huntington′s disease are a direct effect of polyglutamines. Nat. Neurosci. 5, 731–736 (2002).
    Article CAS Google Scholar
  25. Frank, S. et al. The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev. Cell 1, 515–525 (2001).
    Article CAS Google Scholar
  26. Trushina, E. et al. Mutant huntingtin impairs axonal trafficking in mammalian neurons in vivo and in vitro. Mol. Cell. Biol. 24, 8195–8209 (2004).
    Article CAS Google Scholar
  27. Mears, J.A. & Hinshaw, J.E. Visualization of dynamins. Methods Cell Biol. 88, 237–256 (2008).
    Article CAS Google Scholar
  28. Naylor, K. et al. Mdv1 interacts with assembled dnm1 to promote mitochondrial division. J. Biol. Chem. 281, 2177–2183 (2006).
    Article CAS Google Scholar
  29. Neuspiel, M., Zunino, R., Gangaraju, S., Rippstein, P. & McBride, H. Activated mitofusin 2 signals mitochondrial fusion, interferes with Bax activation and reduces susceptibility to radical induced depolarization. J. Biol. Chem. 280, 25060–25070 (2005).
    Article CAS Google Scholar
  30. Jagasia, R., Grote, P., Westermann, B. & Conradt, B. DRP-1–mediated mitochondrial fragmentation during EGL-1–induced cell death in C. elegans. Nature 433, 754–760 (2005).
    Article CAS Google Scholar
  31. Wakabayashi, J. et al. The dynamin-related GTPase Drp1 is required for embryonic and brain development in mice. J. Cell Biol. 186, 805–816 (2009).
    Article CAS Google Scholar
  32. Perkins, G.A. et al. Electron tomographic analysis of cytoskeletal cross-bridges in the paranodal region of the node of Ranvier in peripheral nerves. J. Struct. Biol. 161, 469–480 (2008).
    Article CAS Google Scholar
  33. Ingerman, E. & Nunnari, J. A continuous, regenerative coupled GTPase assay for dynamin-related proteins. Methods Enzymol. 404, 611–619 (2005).
    Article CAS Google Scholar

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Acknowledgements

We thank S. Finkbeiner (University of California–San Francisco) for the pGW1 plasmids encoding huntingtinex1-Q17-GFP, huntingtinex1-Q46-GFP, huntingtinex1-Q97-GFP; L. Thompson (University of California–Irvine) for the huntingtin plasmids pcDNA3.1-Q25-GFP and pcDNA3.1-Q97-GFP; U. Hartl (Max Planck Institute of Biochemistry) for the GST-huntingtinex1-Q20 and -Q53 constructs; A. van der Bliek (University of California–Los Angeles) for the DRP1 K38A cDNA in baculovirus expression vector (US National Center for Biotechnology Information accession number NM_005690.3); R. Youle (US National Institutes of Health (NIH)) for the YFP-DRP1 plasmid; R. Slack (University of Ottawa) for the MFN2 RasG12V (p82-FzoRV12pECFP-C1) expression plasmid; S. Strack (University of Iowa) for his pcDNA3.1β-DRP1shRNA vector; S. Lubitz, J. Johnson, V. DeAssis, C. Eldon and B. Kincaid for technical assistance; and A. Knott for manuscript development and editing. This work is supported by NIH grants to E.B.-W. (R01EY016164 and R01NS055193), a fellowship from the Hereditary Disease Foundation (to G.L.), grants to I.R., M.A.P. and M.R.H. from the Canadian Institutes of Health Research, and support from CHDI to M.R.H. The electron microscope tomography work was carried out in facilities of the US National Center for Microscopy and Imaging Research, supported by NIH grant P41RR004050 awarded to M.E.

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Authors and Affiliations

  1. Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida, USA
    Wenjun Song, Jin Chen, Alejandra Petrilli, Geraldine Liot, Yue Zhou, Blaise Bossy & Ella Bossy-Wetzel
  2. Department of Molecular Biology, Structural Biology Group, University of Salzburg, Salzburg, Austria
    Eva Klinglmayr & Robert Schwarzenbacher
  3. National Center for Microscopy and Imaging Research, University of California–San Diego, San Diego, California, USA
    Patrick Poquiz, Jonathan Tjong, Mark Ellisman & Guy Perkins
  4. Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, British Columbia, Canada
    Mahmoud A Pouladi & Michael R Hayden
  5. University of California–San Diego, La Jolla, California, USA
    Eliezer Masliah
  6. Department of Anatomy and Cell Biology, McGill University, Montreal, Québec, Canada
    Isabelle Rouiller

Authors

  1. Wenjun Song
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  2. Jin Chen
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  3. Alejandra Petrilli
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  4. Geraldine Liot
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  5. Eva Klinglmayr
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  6. Yue Zhou
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  7. Patrick Poquiz
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  8. Jonathan Tjong
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  9. Mahmoud A Pouladi
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  10. Michael R Hayden
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  11. Eliezer Masliah
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  12. Mark Ellisman
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  13. Isabelle Rouiller
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  14. Robert Schwarzenbacher
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  15. Blaise Bossy
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  16. Guy Perkins
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  17. Ella Bossy-Wetzel
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Contributions

W.S. performed all imaging and participated in the mitochondrial fragmentation and cell death analyses. J.C. performed the GTPase assays, some of the immune precipitations and the electron microscopy analysis. A.P. performed some of the neuronal cell death and immune precipitations. G.L. performed the electron microscopy stereology and generated some of the preliminary data. E.K. purified, cloned and prepared the recombinant DRP1 protein. Y.Z. performed western blotting for the DRP1 knockdown. P.P. and J.T. participated in the electron microscope tomography. M.A.P. and M.R.H. provided the YAC18 and YAC128 mice and advice on huntingtin coimmunoprecipitations. E.M. provided human postmortem brain samples. R.S. led the DRP1 protein purification. M.E. and G.P. led the electron microscope tomography experiment. B.B. performed GTPase assays, prepared samples for electron microscopy and purified the huntingtin protein. I.R. performed electron microscope negative-stain experiments. E.B.-W. conceived the project and wrote the article. All authors participated in the data analysis and interpretation of the results.

Corresponding author

Correspondence toElla Bossy-Wetzel.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10, Supplementary Table 1 and Supplementary Methods (PDF 1008 kb)

Supplementary Video 1

Mitochondrial movement in a neuron expressing HTT exon1-Q17-GFP and DsRed2-Mito. Movie corresponds to the kymograph in Figure 1f, top panel and shows mitochondrial transport. The movie lasts 5 min and is played back accelerated (original: 5 s frame−1, playback: 1/6 s frame−1). (MOV 1130 kb)

Supplementary Video 2

Mitochondrial movement in a neuron expressing HTT exon1-Q46-GFP and DsRed2-Mito. Movie corresponds to the kymograph in Figure 1f, center panel and shows a clear decrease in mitochondrial transport. The movie lasts 5 min and is played back accelerated (original: 5 s frame−1, playback: 1/6 s frame−1). (MOV 943 kb)

Supplementary Video 3

Mitochondrial movement in a neuron expressing HTT exon1-Q97-GFP and DsRed2-Mito. Movie corresponds to the kymograph in Figure 1f, bottom panel and shows more pronounced arrest in mitochondrial transport. The movie lasts 5 min and is played back accelerated (original: 5 s frame−1, playback: 1/6 s frame−1). (MOV 607 kb)

Supplementary Video 4

Electron tomography of a control mitochondrion in a medium spiny neuron. Movie showing the three-dimensional details of a mitochondrion in a medium spiny neuron reconstructed using electron tomography. These mitochondria are typically elongated along the direction of the axonal long axis. Clip 1: a rapid sequence through 190 slices (2.2 nm slice−1) of the tomographic volume that shows nearly the entire mitochondrial volume. There are 84 cristae. Clip 2: rotations and zooms of the surface-rendered volume after segmentation of the inner and outer membranes. The blue outer membrane is translucent to visualize the cristae displayed in various colors. Clip 3: rotation of the cristae after removal of the outer membrane to better distinguish the variety of shapes and sizes. (MOV 8195 kb)

Supplementary Video 5

Electron tomography of a fissioning YAC128 mitochondrion in a medium spiny neuron. Movie showing the three-dimensional details of a mitochondrion fissioning into three parts in a medium spiny neuron reconstructed using electron tomography. Clip 1: a rapid sequence through 210 slices (2.2 nm slice−1) of the tomographic volume. There are 223 cristae, many of which are small. Clip 2: rotation showing the outer membrane and the widths of the two constriction sites. Clip 3: rotations showing the cristae in each of the three parts. Clip 4: rotations and zooms highlighting the cristae and the constriction sites. The blue outer membrane is translucent to visualize the cristae displayed in various colors. (MOV 9743 kb)

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Song, W., Chen, J., Petrilli, A. et al. Mutant huntingtin binds the mitochondrial fission GTPase dynamin-related protein-1 and increases its enzymatic activity.Nat Med 17, 377–382 (2011). https://doi.org/10.1038/nm.2313

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