Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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

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.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Mutant huntingtin triggers mitochondrial fragmentation, decreases in anterograde and retrograde transport and neuronal cell death, which depend on polyQ length.
Figure 2: Mutant huntingtin increases the number of small mitochondria and cristae but decreases cristae surface area and volume in the striatum of six-month-old YAC128 mice.
Figure 3: Mutant huntingtin interacts with DRP1 in mice and humans with Huntington's disease and alters DRP1 structure and function.
Figure 4: Restoring mitochondrial fusion with DRP1 K38A or in combination with MFN2 RasG12V rescues neurons from neuritic trafficking defects and cell death.

Accession codes

Accessions

NCBI Reference Sequence

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 

Download references

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.

Author information

Authors and Affiliations

Authors

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 to Ella Bossy-Wetzel.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

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)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2313

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing