Home LiteratureArticle Details
PMID: 19144827 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutant huntingtin impairs post-Golgi trafficking to lysosomes by delocalizing optineurin/Rab8 complex from the Golgi apparatus.

Molecular biology of the cell ·Vol. 20 ·No. 5 ·2009-03-00 ·Pages 1478-92

del Toro D, Alberch J, Lázaro-Diéguez F, Martín-Ibáñez R, Xifró X, Egea G, Canals JM

Abstract

Huntingtin regulates post-Golgi trafficking of secreted proteins. Here, we studied the mechanism by which mutant huntingtin impairs this process. Colocalization studies and Western blot analysis of isolated Golgi membranes showed a reduction of huntingtin in the Golgi apparatus of cells expressing mutant huntingtin. These findings correlated with a decrease in the levels of optineurin and Rab8 in the Golgi apparatus that can be reverted by overexpression of full-length wild-type huntingtin. In addition, immunoprecipitation studies showed reduced interaction between mutant huntingtin and optineurin/Rab8. Cells expressing mutant huntingtin produced both an accumulation of clathrin adaptor complex 1 at the Golgi and an increase of clathrin-coated vesicles in the vicinity of Golgi cisternae as revealed by electron microscopy. Furthermore, inverse fluorescence recovery after photobleaching analysis for lysosomal-associated membrane protein-1 and mannose-6-phosphate receptor showed that the optineurin/Rab8-dependent post-Golgi trafficking to lysosomes was impaired in cells expressing mutant huntingtin or reducing huntingtin levels by small interfering RNA. Accordingly, these cells showed a lower content of cathepsin D in lysosomes, which led to an overall reduction of lysosomal activity. Together, our results indicate that mutant huntingtin perturbs post-Golgi trafficking to lysosomal compartments by delocalizing the optineurin/Rab8 complex, which, in turn, affects the lysosomal function.

MeSH Terms
Animals Cell Cycle Proteins Cell Membrane/metabolism Cells, Cultured Eye Proteins/analysis,metabolism Golgi Apparatus/metabolism Huntingtin Protein Immunohistochemistry Lysosomes/metabolism Membrane Transport Proteins Mice Mutation Nerve Tissue Proteins/analysis,genetics,physiology Nuclear Proteins/analysis,genetics,physiology Protein Transport/physiology Transcription Factor AP-1/metabolism rab GTP-Binding Proteins/analysis,metabolism
Chemicals
Cell Cycle Proteins Eye Proteins Htt protein, mouse Huntingtin Protein Membrane Transport Proteins Nerve Tissue Proteins Nuclear Proteins Optn protein, mouse Rab8 protein, mouse Transcription Factor AP-1 rab GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
del Toro Daniel
Departament de Biologia Cellular, Immunologia i Neurociències, Facultat de Medicina, IDIBAPS, Universitat de Barcelona, E-08036 Barcelona, Spain.
Alberch Jordi
Lázaro-Diéguez Francisco
Martín-Ibáñez Raquel
Xifró Xavier
Egea Gustavo
Canals Josep M
References (39)
39 references, click to expand
  1. Optineurin links myosin VI to the Golgi complex and is involved in Golgi organization and exocytosis.
    J Cell Biol. 2005 Apr 25;169(2):285-95 PMID: 15837803
  2. A worldwide study of the Huntington's disease mutation. The sensitivity and specificity of measuring CAG repeats.
    N Engl J Med. 1994 May 19;330(20):1401-6 PMID: 8159192
  3. Mice deficient for the lysosomal proteinase cathepsin D exhibit progressive atrophy of the intestinal mucosa and profound destruction of lymphoid cells.
    EMBO J. 1995 Aug 1;14(15):3599-608 PMID: 7641679
  4. A kinetic framework for a mammalian RNA polymerase in vivo.
    Science. 2002 Nov 22;298(5598):1623-6 PMID: 12446911
  5. Membrane domains in the secretory and endocytic pathways.
    Cell. 2003 Feb 21;112(4):507-17 PMID: 12600314
  6. FIP-2, a coiled-coil protein, links Huntingtin to Rab8 and modulates cellular morphogenesis.
    Curr Biol. 2000 Dec 14-28;10(24):1603-6 PMID: 11137014
  7. HIP14, a novel ankyrin domain-containing protein, links huntingtin to intracellular trafficking and endocytosis.
    Hum Mol Genet. 2002 Nov 1;11(23):2815-28 PMID: 12393793
  8. Huntingtin expression stimulates endosomal-lysosomal activity, endosome tubulation, and autophagy.
    J Neurosci. 2000 Oct 1;20(19):7268-78 PMID: 11007884
  9. Participation of autophagy in storage of lysosomes in neurons from mouse models of neuronal ceroid-lipofuscinoses (Batten disease).
    Am J Pathol. 2005 Dec;167(6):1713-28 PMID: 16314482
  10. The Rab GTPase family.
    Genome Biol. 2001;2(5):REVIEWS3007 PMID: 11387043
  11. Huntingtin phosphorylation acts as a molecular switch for anterograde/retrograde transport in neurons.
    EMBO J. 2008 Aug 6;27(15):2124-34 PMID: 18615096
  12. Modulation of neuronal protein trafficking and function by palmitoylation.
    Curr Opin Neurobiol. 2005 Oct;15(5):527-35 PMID: 16125924
  13. Huntingtin facilitates dynein/dynactin-mediated vesicle transport.
    Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10045-50 PMID: 17548833
  14. Mannose 6-phosphate receptors and lysosomal enzyme targeting.
    J Biol Chem. 1989 Jul 25;264(21):12115-8 PMID: 2545698
  15. Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles.
    Eur J Cell Biol. 1995 Jan;66(1):3-14 PMID: 7750517
  16. Brain-derived neurotrophic factor regulates the onset and severity of motor dysfunction associated with enkephalinergic neuronal degeneration in Huntington's disease.
    J Neurosci. 2004 Sep 1;24(35):7727-39 PMID: 15342740
  17. Rab8 promotes polarized membrane transport through reorganization of actin and microtubules in fibroblasts.
    J Cell Biol. 1996 Oct;135(1):153-67 PMID: 8858170
  18. The Rab8 GTPase selectively regulates AP-1B-dependent basolateral transport in polarized Madin-Darby canine kidney cells.
    J Cell Biol. 2003 Oct 27;163(2):339-50 PMID: 14581456
  19. Myosin VI is required for sorting of AP-1B-dependent cargo to the basolateral domain in polarized MDCK cells.
    J Cell Biol. 2007 Apr 9;177(1):103-14 PMID: 17403927
  20. Immortalized GABAergic cell lines derived from rat striatum using a temperature-sensitive allele of the SV40 large T antigen.
    Exp Neurol. 1993 Dec;124(2):395-400 PMID: 8287936
  21. The roles of intracellular protein-degradation pathways in neurodegeneration.
    Nature. 2006 Oct 19;443(7113):780-6 PMID: 17051204
  22. Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine.
    Cell. 1984 Dec;39(2 Pt 1):405-16 PMID: 6498939
  23. Imaging proteolysis by living human breast cancer cells.
    Neoplasia. 2000 Nov-Dec;2(6):496-504 PMID: 11228542
  24. The tyrosine-based lysosomal targeting signal in lamp-1 mediates sorting into Golgi-derived clathrin-coated vesicles.
    EMBO J. 1996 Oct 1;15(19):5230-9 PMID: 8895568
  25. Dominant phenotypes produced by the HD mutation in STHdh(Q111) striatal cells.
    Hum Mol Genet. 2000 Nov 22;9(19):2799-809 PMID: 11092756
  26. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules.
    Cell. 2004 Jul 9;118(1):127-38 PMID: 15242649
  27. Widespread expression of Huntington's disease gene (IT15) protein product.
    Neuron. 1995 May;14(5):1065-74 PMID: 7748554
  28. Diacylglycerol is required for the formation of COPI vesicles in the Golgi-to-ER transport pathway.
    Mol Biol Cell. 2007 Sep;18(9):3250-63 PMID: 17567948
  29. Vesicle formation at the plasma membrane and trans-Golgi network: the same but different.
    Science. 2006 Sep 15;313(5793):1591-4 PMID: 16973870
  30. Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease.
    Nat Genet. 2004 Jun;36(6):585-95 PMID: 15146184
  31. Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons.
    Neuron. 1995 May;14(5):1075-81 PMID: 7748555
  32. Small molecule regulators of autophagy identified by an image-based high-throughput screen.
    Proc Natl Acad Sci U S A. 2007 Nov 27;104(48):19023-8 PMID: 18024584
  33. Cathepsin D antigenic epitopes identified by the humoral responses of ovarian cancer patients.
    Cancer Immunol Immunother. 1998 Mar;46(1):48-54 PMID: 9520292
  34. Palmitoylation of huntingtin by HIP14 is essential for its trafficking and function.
    Nat Neurosci. 2006 Jun;9(6):824-31 PMID: 16699508
  35. Mutant huntingtin impairs the post-Golgi trafficking of brain-derived neurotrophic factor but not its Val66Met polymorphism.
    J Neurosci. 2006 Dec 6;26(49):12748-57 PMID: 17151278
  36. Biological basket weaving: formation and function of clathrin-coated vesicles.
    Annu Rev Cell Dev Biol. 2001;17:517-68 PMID: 11687498
  37. Potential therapeutic applications of autophagy.
    Nat Rev Drug Discov. 2007 Apr;6(4):304-12 PMID: 17396135
  38. Perinuclear localization of huntingtin as a consequence of its binding to microtubules through an interaction with beta-tubulin: relevance to Huntington's disease.
    J Cell Sci. 2002 Mar 1;115(Pt 5):941-8 PMID: 11870213
  39. Rab8, a small GTPase involved in vesicular traffic between the TGN and the basolateral plasma membrane.
    J Cell Biol. 1993 Oct;123(1):35-45 PMID: 8408203
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.e08-07-0726
Published
2009-03-00
Epub
2009-00-14
Pages
1478-92
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2649260
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e08-07-0726 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2