Deletions and missense mutations of EPM2A exacerbate unfolded protein response and apoptosis of neuronal cells induced by endoplasm reticulum stress.
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.
Liu
Yan
Y
Division of Immunotherapy, Section of General Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI 48109, USA.
Wang
Yin
Y
Wu
Cindy
C
Liu
Yang
Y
Zheng
Pan
P
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020192:Lafora Disease / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D051379:Mice; D009154:Mutation; D020125:Mutation, Missense; D041681:NIH 3T3 Cells; D009474:Neurons / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D017510:Protein Folding; D054558:Protein Tyrosine Phosphatases, Non-Receptor / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion
D054558:Protein Tyrosine Phosphatases, Non-Receptor; C114990:EPM2A protein, human