Human Molecular Genetics Advance Access originally published online on April 29, 2009
Human Molecular Genetics 2009 18(14):2622-2631; doi:10.1093/hmg/ddp196
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Deletions and missense mutations of EPM2A exacerbate unfolded protein response and apoptosis of neuronal cells induced by endoplasm reticulum stress
1 Division of Immunotherapy, Section of General Surgery, Department of Surgery 2 Department of Internal Medicine 3 Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA 4 Department of Pathology, The Ohio State University Medical Center, Columbus, OH 43210, USA
* To whom correspondence should be addressed at: Department of Surgery and Pathology, University of Michigan, BSRB 1810, 109 Zina Pitcher Place, Ann Arbor, MI 48109. Tel: +1 7346153464; Fax: +1 7347632162; Email: panz{at}umich.edu
Received March 9, 2009; Accepted April 27, 2009
The majority of the Lafora's disease (LD) is caused by defect in the EPM2A gene, including missense and nonsense mutations and deletions. These defects mainly occur in the carbohydrate-binding domain, and how these mutations cause neuronal defects is under active investigation. Here, we report that the mutant proteins encoded by all missense mutations and most deletions tested are unstable, insoluble and ubiquitinated, and are accumulated in aggresome-like structures. The effect of apparent ‘gain-of-function’ mutations can be corrected by co-transfection of wild-type EPM2A cDNA, which is consistent with the recessive nature of these mutations in LD patients. In a neuronal cell line, these mutant aggregates exacerbate endoplasm reticulum (ER) stress and make the cells susceptible to the apoptosis induced by ER stressor, thapsigargin. The chemical chaperon, 4-phenylbutyrate, increased the mutant solubility, reduced the ER stress and dulled the sensitivity of mutant neuronal cells to apoptosis induced by thapsigargin and the mutant laforin proteins. The increased sensitivity to ER stress-induced apoptosis may contribute to LD pathogenesis.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.