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PMID: 15684030 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

The vacuolar kinase Yck3 maintains organelle fragmentation by regulating the HOPS tethering complex.

The Journal of cell biology ·Vol. 168 ·No. 3 ·2005-01-31 ·Pages 401-14

LaGrassa TJ, Ungermann C

Abstract

The regulation of cellular membrane flux is poorly understood. Yeast respond to hypertonic stress by fragmentation of the normally large, low copy vacuole. We used this phenomenon as the basis for an in vivo screen to identify regulators of vacuole membrane dynamics. We report here that maintenance of the fragmented phenotype requires the vacuolar casein kinase I Yck3: when Yck3 is absent, salt-stressed vacuoles undergo fission, but reassemble in a SNARE-dependent manner, suggesting that vacuole fusion is disregulated. Accordingly, when Yck3 is deleted, in vitro vacuole fusion is increased, and Yck3 overexpression blocks fusion. Morphological and functional studies show that Yck3 modulates the Rab/homotypic fusion and vacuole protein sorting complex (HOPS)-dependent tethering stage of vacuole fusion. Intriguingly, Yck3 mediates phosphorylation of the HOPS subunit Vps41, a bi-functional protein involved in both budding and fusion during vacuole biogenesis. Because Yck3 also promotes efficient vacuole inheritance, we propose that tethering complex phosphorylation is a part of a general, switch-like mechanism for driving changes in organelle architecture.

MeSH Terms
Adaptor Proteins, Vesicular Transport Antibodies/pharmacology Casein Kinase I/genetics,metabolism,physiology Enzyme Inhibitors/pharmacology Genotype Guanine Nucleotide Dissociation Inhibitors/pharmacology Marine Toxins Membrane Fusion/drug effects,physiology Membrane Proteins/immunology,metabolism Microcystins Microscopy, Fluorescence Models, Biological Mutation Nerve Tissue Proteins/immunology Organelles/metabolism,physiology Peptides, Cyclic/pharmacology Phosphorylation Phosphotransferases (Alcohol Group Acceptor)/genetics SNARE Proteins Saccharomyces cerevisiae/drug effects,genetics,physiology Saccharomyces cerevisiae Proteins/agonists,antagonists & inhibitors,genetics,immunology,metabolism,physiology Saline Solution, Hypertonic/pharmacology Synaptosomal-Associated Protein 25 Vacuoles/metabolism,physiology Vesicular Transport Proteins/genetics,immunology,metabolism,physiology rab GTP-Binding Proteins/antagonists & inhibitors,immunology ras GTPase-Activating Proteins/agonists
Chemicals
Adaptor Proteins, Vesicular Transport Antibodies Enzyme Inhibitors GDP dissociation inhibitor 1 Guanine Nucleotide Dissociation Inhibitors Gyp7 protein, S cerevisiae Marine Toxins Membrane Proteins Microcystins Nerve Tissue Proteins Peptides, Cyclic SNARE Proteins Saccharomyces cerevisiae Proteins Saline Solution, Hypertonic Synaptosomal-Associated Protein 25 VAM6 protein, S cerevisiae VPS3 protein, S cerevisiae VPS41 protein, S cerevisiae Vesicular Transport Proteins ras GTPase-Activating Proteins FAB1 protein, S cerevisiae Phosphotransferases (Alcohol Group Acceptor) Casein Kinase I YCK3 protein, S cerevisiae YPT7 protein, S cerevisiae rab GTP-Binding Proteins cyanoginosin LR
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
LaGrassa Tracy J
Biochemie-Zentrum der Universität Heidelberg, 69120 Heidelberg, Germany.
Ungermann Christian
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200407141
Published
2005-01-31
Pages
401-14
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2171739
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.200407141 PMC Full Text View journal details
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