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Modular Dispensability of Dysferlin C2 Domains Reveals Rational Design for Mini-dysferlin Molecules

2012· article· en· 26 citations· W2027513958 on OpenAlex· 10.1074/jbc.m112.391722

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Canadian affiliationAn author listed a Canadian institution. This is the only route the usual frame has.

Post-publication record

Nature
Retraction
Reason
Duplication of/in Image;Falsification/Fabrication of Data;Investigation by Company/Institution;
Date
7/28/2017 0:00
Flagged by OpenAlex?
Yes

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Abstract

Dysferlin is a large transmembrane protein composed of a C-terminal transmembrane domain, two DysF domains, and seven C2 domains that mediate lipid- and protein-binding interactions. Recessive loss-of-function mutations in dysferlin lead to muscular dystrophies, for which no treatment is currently available. The large size of dysferlin precludes its encapsulation into an adeno-associated virus (AAV), the vector of choice for gene delivery to muscle. To design mini-dysferlin molecules suitable for AAV-mediated gene transfer, we tested internally truncated dysferlin constructs, each lacking one of the seven C2 domains, for their ability to localize to the plasma membrane and to repair laser-induced plasmalemmal wounds in dysferlin-deficient human myoblasts. We demonstrate that the dysferlin C2B, C2C, C2D, and C2E domains are dispensable for correct plasmalemmal localization. Furthermore, we show that the C2B, C2C, and C2E domains and, to a lesser extent, the C2D domain are dispensable for dysferlin membrane repair function. On the basis of these results, we designed small dysferlin molecules that can localize to the plasma membrane and reseal laser-induced plasmalemmal injuries and that are small enough to be incorporated into AAV. These results lay the groundwork for AAV-mediated gene therapy experiments in dysferlin-deficient mouse models.

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The record

Venue
Journal of Biological Chemistry
Topic
Muscle Physiology and Disorders
Field
Biochemistry, Genetics and Molecular Biology
Canadian institutions
McGill UniversityMontreal Neurological Institute and Hospital
Funders
Keywords
DysferlinTransmembrane domainTransmembrane proteinCell biologyBiologyChemistryMuscular dystrophyGeneGeneticsReceptor
Has abstract in OpenAlex
yes