Epileptic encephalopathy-causing mutations in DNM1 impair synaptic vesicle endocytosis.

ABSTRACT:

Objective

To elucidate the functional consequences of epileptic encephalopathy-causing de novo mutations in DNM1 (A177P, K206N, G359A), which encodes a large mechanochemical GTPase essential for neuronal synaptic vesicle endocytosis.

Methods

HeLa and COS-7 cells transfected with wild-type and mutant DNM1 constructs were used for transferrin assays, high-content imaging, colocalization studies, Western blotting, and electron microscopy (EM). EM was also conducted on the brain sections of mice harboring a middle-domain Dnm1 mutation (Dnm1 (Ftfl)).

Results

We demonstrate that the expression of each mutant protein decreased endocytosis activity in a dominant-negative manner. One of the G-domain mutations, K206N, decreased protein levels. The G359A mutation, which occurs i

SUBMITTER: Dhindsa RS 

PROVIDER: S-EPMC4821085 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature