title: Insights into dynamin-associated disorders through analysis of equivalent mutations in the yeast dynamin Vps1.
abstract: The dynamins represent a superfamily of proteins that have been shown to function in a wide range of membrane fusion and fission events. An increasing number of mutations in the human classical dynamins, Dyn-1 and Dyn-2 has been reported, with diseases caused by these changes ranging from Charcot-Marie-Tooth disorder to epileptic encephalopathies. The budding yeast,Saccharomyces cerevisiaeexpresses a single dynamin-related protein that functions in membrane trafficking, and is considered to play a similar role to Dyn-1 and Dyn-2 during scission of endocytic vesicles at the plasma membrane. Large parts of the dynamin protein are highly conserved across species and this has enabled us in this study to select a number of disease causing mutations and to generate equivalent mutations in Vps1. We have then studied these mutants using both cellular and biochemical assays to ascertain functions of the protein that have been affected by the changes. Specifically, we demonstrate that the Vps1-G397R mutation (Dyn-2 G358R) disrupts protein oligomerization, Vps1-A447T (Dyn-1 A408T) affects the scission stage of endocytosis, while Vps1-R298L (Dyn-1 R256L) affects lipid binding specificity and possibly an early stage in endocytosis. Overall, we consider that the yeast model will potentially provide an avenue for rapid analysis of new dynamin mutations in order to understand the underlying mechanisms that they disrupt.
authors.affiliation: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Laila
authors.initials: L
authors.lastname: Moustaq
authors.affiliation: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Iwona I
authors.initials: II
authors.lastname: Smaczynska-de Rooij
authors.affiliation: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Palmer
authors.affiliation: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Marklew
authors.affiliation: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors.forename: Kathryn R
authors.initials: KR
authors.lastname: Ayscough
keywords: Charcot-Marie-Tooth; Disease mutation; Dynamin; Epilepsy; Saccharomyces cerevisiae