Microtubule disruption inhibits autophagosome-lysosome fusion: implications for studying the roles of aggresomes in polyglutamine diseases.
Large cytoplasmic inclusions called aggresomes are seen in many protein conformational diseases including Huntington's disease and Parkinson's disease. The roles of inclusions and aggresomes in these diseases are unresolved critical issues that have been vigorously debated. Two recent studies used microtubule disruption with nocodazole to inhibit aggresome formation and observed increased toxicity of expanded polyglutamines in the context of huntingtin exon 1 and a truncated androgen receptor. Increased toxicity of expanded polyglutamines in the presence of nocodazole was correlated with decreased protein turnover, leading the authors to conclude that aggresomes were cytoprotective and that they directly enhanced clearance of the toxic proteins. Here we show that nocodazole has additional effects, which provide a simple alternative explanation for these previous observations. We confirmed aggresome formation in cells expressing proteins with polyalanine and polyglutamine expansions. As expected, we found a reduction in aggresome formation when microtubule function was disrupted using nocodazole. However, in addition to this effect, nocodazole treatment increased the proportions of cells with nuclear inclusions in PC12 cells expressing huntingtin exon 1 with 74 glutamines. This can be explained as nocodazole inhibits autophagosome-lysosome fusion, a key step in mutant huntingtin exon 1 clearance. This effect alone can explain the previous observations with this compound in polyglutamine diseases and raises doubts about the interpretation of some of the data that have been used to argue that aggresomes protect against polyglutamine mutations.
Department of Medical Genetics, Cambridge Institute for Medical Research, Wellcome/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2XY, UK.
Julie L
JL
Webb
Brinda
B
Ravikumar
David C
DC
Rubinsztein
D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D006801:Humans; D006816:Huntington Disease / Q000503:physiopathology; D002479:Inclusion Bodies / Q000502:physiology*; D008247:Lysosomes / Q000187:drug effects / Q000502:physiology*; D008561:Membrane Fusion; D008870:Microtubules / Q000187:drug effects / Q000502:physiology*; D019636:Neurodegenerative Diseases / Q000503:physiopathology*; D015739:Nocodazole / Q000494:pharmacology; D030781:Organisms, Genetically Modified; D016716:PC12 Cells; D010300:Parkinson Disease / Q000503:physiopathology; D010455:Peptides / Q000502:physiology*; D051381:Rats
D010455:Peptides; C019529:polyalanine; C097188:polyglutamine; D015739:Nocodazole