Laforin preferentially binds the neurotoxic starch-like polyglucosans, which form in its absence in progressive myoclonus epilepsy.
Lafora disease (LD) is a fatal and the most common form of adolescent-onset progressive epilepsy. Fulminant endoplasmic reticulum (ER)-associated depositions of starch-like long-stranded, poorly branched glycogen molecules [known as polyglucosans, which accumulate to form Lafora bodies (LBs)] are seen in neuronal perikarya and dendrites, liver, skeletal muscle and heart. The disease is caused by loss of function of the laforin dual-specificity phosphatase or the malin E3 ubiquitin ligase. Towards understanding the pathogenesis of polyglucosans in LD, we generated a transgenic mouse overexpressing inactivated laforin to trap normal laforin's unknown substrate. The trap was successful and LBs formed in liver, muscle, neuronal perikarya and dendrites. Using immunogold electron microscopy, we show that laforin is found in close proximity to the ER surrounding the polyglucosan accumulations. In neurons, it compartmentalizes to perikaryon and dendrites and not to axons. Importantly, it binds polyglucosans, establishing for the first time a direct association between the disease-defining storage product and disease protein. It preferentially binds polyglucosans over glycogen in vivo and starch over glycogen in vitro, suggesting that laforin's role begins after the appearance of polyglucosans and that the laforin pathway is involved in monitoring for and then preventing the formation of polyglucosans. In addition, we show that the laforin interacting protein, EPM2AIP1, also localizes on the polyglucosan masses, and we confirm laforin's intense binding to LBs in human LD biopsy material.
Chan
Elayne M
EM
The Hospital for Sick Children and Department of Molecular and Medical Genetics, the University of Toronto, Toronto, Canada.
Ackerley
Cameron A
CA
Lohi
Hannes
H
Ianzano
Leonarda
L
Cortez
Miguel A
MA
Shannon
Patrick
P
Scherer
Stephen W
SW
Minassian
Berge A
BA
D000199:Actins / Q000235:genetics; D000818:Animals; D001921:Brain / Q000473:pathology; D054637:Dual-Specificity Phosphatases; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D018014:Gene Transfer Techniques; D005936:Glucans / Q000378:metabolism*; D006003:Glycogen / Q000378:metabolism; D006801:Humans; D007120:Immunochemistry; D002479:Inclusion Bodies / Q000648:ultrastructure; D020192:Lafora Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008099:Liver / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D054558:Protein Tyrosine Phosphatases, Non-Receptor; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D013213:Starch / Q000378:metabolism
D000199:Actins; D005936:Glucans; C489427:MYC protein, human; C489428:Myc protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D013213:Starch; D006003:Glycogen; C083094:polyglucosan; D054637:Dual-Specificity Phosphatases; C491989:Epm2a protein, mouse; D017027:Protein Tyrosine Phosphatases; D054558:Protein Tyrosine Phosphatases, Non-Receptor; C114990:EPM2A protein, human