Search Organism: All OrganismsArabidopsis thalianaBacillus subtilis 168Bos taurusCaenorhabditis elegansCanis familiarisDanio rerioDrosophila melanogasterEquus caballusEscherichia coli K12Gallus gallusHomo sapiensMacaca mulattaMus musculusNematostella vectensisOryza sativaPan troglodytesRattus norvegicusSaccharomyces cerevisiaeSchizosaccharomyces pombeStrongylocentrotus purpuratusTakifugu rubripesXenopus laevis home help / support contribute downloads mirrors about us Traffic Dec (2007); 8(12):1829-40 Dominant-Negative Behavior of Mammalian Vps35 in Yeast Requires a Conserved PRLYL Motif Involved in Retromer Assembly- Zhao X, Nothwehr S, Lara-Lemus R, Zhang BY, Peter H, Arvan P Division of Metabolism, Endocrinology - Diabetes, University of Michigan, 5560 MSRB2, 1150 W- Medical Center Drive, Ann Arbor, MI 48109, USA- Abstract: The retromer protein complex assists in recycling selected integral membrane proteins from endosomes to the trans Golgi network- One protein subcomplex -Vps35p, Vps26p and Vps29p- combines with a second -Vps17p and Vps5p- to form a coat involved in sorting and budding of endosomal vesicles- Yeast Vps35p -yVps35- exhibits similarity to human Vps35 -hVps35-, especially in a completely conserved PRLYL motif contained within an amino-terminal domain- Companion studies indicate that an R-98-W mutation in yVps35 causes defective retromer assembly in Saccharomyces cerevisiae- Herein, we find that the expression of hVps35 in yeast confers dominant-negative vacuolar proenzyme secretion and defective secretory proprotein processing- The mutant phenotype appears to be driven by hVps35 competing with endogenous yVps35, becoming incorporated into defective retromer complexes and causing proteasomal degradation of endogenous Vps26 and Vps29- Increased expression of yVps35 displaces some hVps35 to a 100 000 x g supernatant and suppresses the dominant-negative phenotype- Remarkably, mutation of the conserved R-107-W of hVps35 displaces some of the protein to the 100 000 x g supernatant, slows protein turnover and restores stability of Vps26p and Vps29p and completely abrogates dominant-negative trafficking behavior- We show that hVps35 coprecipitates Vps26, whereas the R-107-W mutant does not- In pancreatic beta cells, the R-107-W mutant shifts hVps35 from peripheral endosomes to a juxtanuclear compartment, affecting both mannose phosphate receptors and insulin- These data underscore importance of the Vps35 PRLYL motif in retromer subcomplex interactions and function- [PUBMED: 17916227] Copyright © 2009, Tyers Lab, The Samuel Lunenfeld Research Institute, Mount Sinai Hospital, All Rights Reserved. terms and conditions - privacy policy - Osprey Network Visualization System BioGRID: A General Repository for Interaction Datasets. Chris Stark, Bobby-Joe Breitkreutz, Teresa Reguly, Lorrie Boucher, Ashton Breitkreutz, Mike Tyers. Nucleic Acids Res. Jan 1;34:D535-9.