Systematic discovery of Rab GTPases with synaptic functions in Drosophila

Date
2011
Authors
Chan, C.-C.
Scoggin, S.
Wang, D.
Cherry, S.
Dembo, T.
Greenberg, B.
Jin, E.J.
Kuey, C.
Lopez, A.
Mehta, S.Q.
Advisor
Instructor
Source Title
Current Biology
Print ISSN
0960-9822
Electronic ISSN
Publisher
Volume
21
Issue
20
Pages
1704 - 1715
Language
English
Type
Article
Journal Title
Journal ISSN
Volume Title
Abstract

Background: Neurons require highly specialized intracellular membrane trafficking, especially at synapses. Rab GTPases are considered master regulators of membrane trafficking in all cells, and only very few Rabs have known neuron-specific functions. Here, we present the first systematic characterization of neuronal expression, subcellular localization, and function of Rab GTPases in an organism with a brain. Results: We report the surprising discovery that half of all Drosophila Rabs function specifically or predominantly in distinct subsets of neurons in the brain. Furthermore, functional profiling of the GTP/GDP-bound states reveals that these neuronal Rabs are almost exclusively active at synapses and the majority of these synaptic Rabs specifically mark synaptic recycling endosomal compartments. Our profiling strategy is based on Gal4 knockins in large genomic fragments that are additionally designed to generate mutants by ends-out homologous recombination. We generated 36 large genomic targeting vectors and transgenic rab-Gal4 fly strains for 25 rab genes. Proof-of-principle knockout of the synaptic rab27 reveals a sleep phenotype that matches its cell-specific expression. Conclusions: Our findings suggest that up to half of all Drosophila Rabs exert specialized synaptic functions. The tools presented here allow systematic functional studies of these Rabs and provide a method that is applicable to any large gene family in Drosophila. © 2011 Elsevier Ltd. All rights reserved.

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Keywords
Drosophila protein, Rab protein, Rab27 protein, Drosophila, animal, antibody specificity, article, bacterial artificial chromosome, brain, Drosophila, gene expression regulation, gene inactivation, genetics, homologous recombination, metabolism, multigene family, mutation, nerve cell, phenotype, physiology, synapse, transgenic animal, Animals, Animals, Genetically Modified, Brain, Chromosomes, Artificial, Bacterial, Drosophila, Drosophila Proteins, Gene Expression Regulation, Enzymologic, Gene Knockout Techniques, Homologous Recombination, Multigene Family, Mutation, Neurons, Organ Specificity, Phenotype, rab GTP-Binding Proteins, Synapses
Citation
Published Version (Please cite this version)