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Palmitoylation of huntingtin by HIP14is essential for its trafficking and function

Nature Neuroscience volume 9, pages 824–831 (2006)Cite this article

Abstract

Post-translational modification by the lipid palmitate is crucial for the correct targeting and function of many proteins. Here we show that huntingtin (htt) is normally palmitoylated at cysteine 214, which is essential for its trafficking and function. The palmitoylation and distribution of htt are regulated by the palmitoyl transferase huntingtin interacting protein 14 (HIP14). Expansion of the polyglutamine tract of htt, which causes Huntington disease, results in reduced interaction between mutant htt and HIP14 and consequently in a marked reduction in palmitoylation. Mutation of the palmitoylation site of htt, making it palmitoylation resistant, accelerates inclusion formation and increases neuronal toxicity. Downregulation of HIP14 in mouse neurons expressing wild-type and mutant htt increases inclusion formation, whereas overexpression of HIP14 substantially reduces inclusions. These results suggest that the expansion of the polyglutamine tract in htt results in decreased palmitoylation, which contributes to the formation of inclusion bodies and enhanced neuronal toxicity.

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Figure 1: Huntingtin is palmitoylated in neurons and COS cells.
Figure 2: Increased inclusions of palmitoylation-resistant mutant htt in COS cells.
Figure 3: Altered distribution of palmitoylation-resistant mutant htt in COS cells and neurons.
Figure 4: Enhanced toxicity of palmitoylation-resistant mutant htt.
Figure 5: HIP14 influences the distribution and catalyzes the palmitoylation of htt in neurons.
Figure 6: HIP14 associates less with mutant htt, and mutant htt is less palmitoylated in vivo.
Figure 7: HIP14 regulates the palmitoylation and distribution of huntingtin in vivo.

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References

  1. El-Husseini, A.E. et al. Dual palmitoylation of PSD-95 mediates its vesiculotubular sorting, postsynaptic targeting, and ion channel clustering. J. Cell Biol. 148, 159–172 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. El-Husseini, A. & Bredt, D.S. Protein palmitoylation: a regulator of neuronal development and function. Nat. Rev. Neurosci. 3, 791–802 (2002).

    Article  CAS  Google Scholar 

  3. Huang, K. & El-Husseini, A. Modulation of neuronal protein trafficking and function by palmitoylation. Curr. Opin. Neurobiol. 15, 527–535 (2005).

    Article  CAS  PubMed  Google Scholar 

  4. Huang, K. et al. Huntingtin-interacting protein HIP14 is a palmitoyl transferase involved in palmitoylation and trafficking of multiple neuronal proteins. Neuron 44, 977–986 (2004).

    Article  CAS  PubMed  Google Scholar 

  5. The Huntington's Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. Cell 72, 971–983 (1993).

  6. Hackam, A.S. et al. The influence of huntingtin protein size on nuclear localization and cellular toxicity. J. Cell Biol. 141, 1097–1105 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Wheeler, V.C. et al. Early phenotypes that presage late-onset neurodegenerative disease allow testing of modifiers in Hdh CAG knock-in mice. Hum. Mol. Genet. 11, 633–640 (2002).

    Article  CAS  PubMed  Google Scholar 

  8. Van Raamsdonk, J.M., Murphy, Z., Slow, E.J., Leavitt, B.R. & Hayden, M.R. Selective degeneration and nuclear localization of mutant huntingtin in the YAC128 mouse model of Huntington disease. Hum. Mol. Genet. 14, 3823–3835 (2005).

    Article  CAS  PubMed  Google Scholar 

  9. Velier, J. et al. Wild-type and mutant huntingtins function in vesicle trafficking in the secretory and endocytic pathways. Exp. Neurol. 152, 34–40 (1998).

    Article  CAS  PubMed  Google Scholar 

  10. DiFiglia, M. et al. Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron 14, 1075–1081 (1995).

    Article  CAS  PubMed  Google Scholar 

  11. Kegel, K.B. et al. Huntingtin associates with acidic phospholipids at the plasma membrane. J. Biol. Chem. 280, 36464–36473 (2005).

    Article  CAS  PubMed  Google Scholar 

  12. Kalchman, M.A. et al. Huntingtin is ubiquitinated and interacts with a specific ubiquitin-conjugating enzyme. J. Biol. Chem. 271, 19385–19394 (1996).

    Article  CAS  PubMed  Google Scholar 

  13. Steffan, J.S. et al. SUMO modification of Huntingtin and Huntington's disease pathology. Science 304, 100–104 (2004).

    Article  CAS  PubMed  Google Scholar 

  14. Humbert, S. et al. The IGF-1/Akt pathway is neuroprotective in Huntington's disease and involves Huntingtin phosphorylation by Akt. Dev. Cell 2, 831–837 (2002).

    Article  CAS  PubMed  Google Scholar 

  15. Warby, S.C. et al. Huntingtin phosphorylation on serine 421 is significantly reduced in the striatum and by polyglutamine expansion in vivo. Hum. Mol. Genet. 14, 1569–1577 (2005).

    Article  CAS  PubMed  Google Scholar 

  16. Kopito, R.R. Aggresomes, inclusion bodies and protein aggregation. Trends Cell Biol. 10, 524–530 (2000).

    Article  CAS  PubMed  Google Scholar 

  17. Waelter, S. et al. Accumulation of mutant huntingtin fragments in aggresome-like inclusion bodies as a result of insufficient protein degradation. Mol. Biol. Cell 12, 1393–1407 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Wyttenbach, A. et al. Heat shock protein 27 prevents cellular polyglutamine toxicity and suppresses the increase of reactive oxygen species caused by huntingtin. Hum. Mol. Genet. 11, 1137–1151 (2002).

    Article  CAS  PubMed  Google Scholar 

  19. Sapp, E. et al. Huntingtin localization in brains of normal and Huntington's disease patients. Ann. Neurol. 42, 604–612 (1997).

    Article  CAS  PubMed  Google Scholar 

  20. Zeron, M.M. et al. Increased sensitivity to N-methyl-D-aspartate receptor-mediated excitotoxicity in a mouse model of Huntington's disease. Neuron 33, 849–860 (2002).

    Article  CAS  PubMed  Google Scholar 

  21. Singaraja, R.R. et al. HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis. Hum. Mol. Genet. 11, 2815–2828 (2002).

    Article  CAS  PubMed  Google Scholar 

  22. 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  PubMed  Google Scholar 

  23. Drisdel, R.C. & Green, W.N. Labeling and quantifying sites of protein palmitoylation. Biotechniques 36, 276–285 (2004).

    Article  CAS  PubMed  Google Scholar 

  24. Drisdel, R.C., Manzana, E. & Green, W.N. The role of palmitoylation in functional expression of nicotinic alpha7 receptors. J. Neurosci. 24, 10502–10510 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. El-Husseini, A. et al. Synaptic strength regulated by palmitate cycling on PSD-95. Cell 108, 849–863 (2002).

    Article  CAS  Google Scholar 

  26. Rakhilin, S. et al. Alpha-bungarotoxin receptors contain alpha7 subunits in two different disulfide-bonded conformations. J. Cell Biol. 146, 203–218 (1999).

    CAS  PubMed Central  PubMed  Google Scholar 

  27. Rocks, O. et al. An acylation cycle regulates localization and activity of palmitoylated Ras isoforms. Science 307, 1746–1752 (2005).

    Article  CAS  PubMed  Google Scholar 

  28. Drenan, R.M. et al. Palmitoylation regulates plasma membrane-nuclear shuttling of R7BP, a novel membrane anchor for the RGS7 family. J. Cell Biol. 169, 623–633 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Ferrante, R.J. et al. Histone deacetylase inhibition by sodium butyrate chemotherapy ameliorates the neurodegenerative phenotype in Huntington's disease mice. J. Neurosci. 23, 9418–9427 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Mastroberardino, P.G. et al. 'Tissue' transglutaminase ablation reduces neuronal death and prolongs survival in a mouse model of Huntington's disease. Cell Death Differ. 9, 873–880 (2002).

    Article  CAS  PubMed  Google Scholar 

  31. Slow, E.J. et al. Absence of behavioral abnormalities and neurodegeneration in vivo despite widespread neuronal huntingtin inclusions. Proc. Natl. Acad. Sci. USA 102, 11402–11407 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Arrasate, M., Mitra, S., Schweitzer, E.S., Segal, M.R. & Finkbeiner, S. Inclusion body formation reduces levels of mutant huntingtin and the risk of neuronal death. Nature 431, 805–810 (2004).

    Article  CAS  PubMed  Google Scholar 

  33. Soyombo, A.A. & Hofmann, S.L. Molecular cloning and expression of palmitoyl-protein thioesterase 2 (PPT2), a homolog of lysosomal palmitoyl-protein thioesterase with a distinct substrate specificity. J. Biol. Chem. 272, 27456–27463 (1997).

    Article  CAS  PubMed  Google Scholar 

  34. Verkruyse, L.A. & Hofmann, S.L. Lysosomal targeting of palmitoyl-protein thioesterase. J. Biol. Chem. 271, 15831–15836 (1996).

    Article  CAS  PubMed  Google Scholar 

  35. Wellington, C.L. et al. Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J. Biol. Chem. 273, 9158–9167 (1998).

    Article  CAS  PubMed  Google Scholar 

  36. Janas, J., Skowronski, J. & Van, A.L. Lentiviral delivery of RNAi in hippocampal neurons. Methods Enzymol. 406, 593–605 (2006).

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank O. Sadiq and E. Eyu for technical assistance. M.R.H. is supported by grants from the Canadian Institutes for Health Research, the Huntington Disease Society of America, the Jack and Doris Brown Foundation and the Huntington Society of Canada. A.E.H. is supported by grants from the Canadian Institutes for Health Research, the EJLB foundation and Neuroscience Canada. R.C.D. and W.N.G. are supported by funding from the National Institute of Neurological Diseases and Stroke, the National Institute of Drug Abuse and the Alzheimer's Association. D.C.R. is funded by a Wellcome Trust Senior Fellowship in Clinical Science, an M.R.C. programme grant and E.U. Framework VI (EUROSCA). A.Y. is supported by funding from the Michael Smith Foundation for Health Research. KH is supported by University Graduate Fellowship and HighQ. M.R.H. and A.E.H. are supported by funding from the HighQ foundation and are investigators of the Fundamental Innovation in Neurodegenerative Diseases (FIND) Research Infrastructure Unit, funded by the Michael Smith Foundation for Health Research. M.R.H. holds a Canada Research Chair in Human Genetics and is a University Killam Professor.

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Author notes

  1. Anat Yanai, Kun Huang and Rujun Kang: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, V5Z 4H4, British Columbia, Canada

    Anat Yanai, Roshni R Singaraja, Lu Gan, Paul C Orban & Michael R Hayden

  2. Department of Psychiatry, University of British Columbia, Vancouver, V6T 1Z3, British Columbia, Canada

    Kun Huang, Rujun Kang, Pamela Arstikaitis, Asher Mullard, Catherine M Cowan, Lynn A Raymond & Alaa El-Husseini

  3. Department of Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, 60637, Illinois, USA

    Renaldo C Drisdel & William N Green

  4. Department of Medical Genetics, Cambridge Institute for Medical Research, Wellcome/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2XY, UK

    Brinda Ravikumar & David C Rubinsztein

  5. Children and Family Research Institute, British Columbia Children's Hospital, Vancouver, V5Z 4H4, British Columbia, Canada

    Michael R Hayden

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Contributions

A.Y. performed all DNA manipulations to generate truncated and full-length mutant htt and HIP14 proteins, and performed most of the [3H]palmitoylation assays and data analyses. K.H. conducted all the Btn-BMCC palmitoylation assays for full-length htt, scored cells for inclusions in htt-transfected COS cells and neurons, designed and characterized HIP14 siRNA, performed all experiments (and corresponding data analysis) in which the alteration of htt trafficking was investigated by knocking down HIP14, conducted the virus production and infection experiments, and scored for TUNEL assay. R.K. performed endogenous htt [3H]palmitoylation assay, scored inclusions in neurons transfected with full-length htt, and performed fragment-htt toxicity assay and the corresponding data analysis. R.R.S. and L.G. performed the htt and HIP14 coimmunoprecipitation experiment. P.C.O. generated HIP14 siRNA lentiviral constructs and supervised the virus production experiment. P.A. conducted the time-lapse experiments. A.M. assisted in the characterization of full-length htt palmitoylation. C.M.C. and L.A.R. were involved in developing the NMDA-induced excitotoxicity TUNEL assay. R.K. and K.H. conducted the TUNEL assays. W.N.G. and R.C.D. developed and supervised the Btn-BMCC palmitoylation assays. B.R. and D.C.R. performed toxicity assays in COS cells. M.R.H., A.E.H., A.Y. and K.H. wrote the manuscript. M.R.H. and A.E.H. supervised the project.

Corresponding authors

Correspondence to Alaa El-Husseini or Michael R Hayden.

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Supplementary information

Supplementary Fig. 1 (PDF 1256 kb)

Supplementary Fig. 2

Altered distribution of palmitoylation-resistant htt in HEK-293 cells. (PDF 1609 kb)

Supplementary Fig. 3 (PDF 2603 kb)

Supplementary Fig. 4 (PDF 4040 kb)

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Yanai, A., Huang, K., Kang, R. et al. Palmitoylation of huntingtin by HIP14is essential for its trafficking and function. Nat Neurosci 9, 824–831 (2006). https://doi.org/10.1038/nn1702

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