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PMID: 10888679 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Luv1p/Rki1p/Tcs3p/Vps54p, a yeast protein that localizes to the late Golgi and early endosome, is required for normal vacuolar morphology.

Molecular biology of the cell ·Vol. 11 ·No. 7 ·2000-07-00 ·Pages 2429-43

Conboy MJ, Cyert MS

Abstract

We have characterized LUV1/RKI1/TCS3/VPS54, a novel yeast gene required to maintain normal vacuolar morphology. The luv1 mutant was identified in a genetic screen for mutants requiring the phosphatase calcineurin for vegetative growth. luv1 mutants lack a morphologically intact vacuole and instead accumulate small vesicles that are acidified and contain the vacuolar proteins alkaline phosphatase and carboxypeptidase Y and the vacuolar membrane H(+)-ATPase. Endocytosis appears qualitatively normal in luv1 mutants, but some portion (28%) of carboxypeptidase Y is secreted. luv1 mutants are sensitive to several ions (Zn(2+), Mn(2+), and Cd(2+)) and to pH extremes. These mutants are also sensitive to hygromycin B, caffeine, and FK506, a specific inhibitor of calcineurin. Some vacuolar protein-sorting mutants display similar drug and ion sensitivities, including sensitivity to FK506. Luv1p sediments at 100,000 x g and can be solubilized by salt or carbonate, indicating that it is a peripheral membrane protein. A Green Fluorescent Protein-Luv1 fusion protein colocalizes with the dye FM 4-64 at the endosome, and hemagglutinin-tagged Luv1p colocalizes with the trans-Golgi network/endosomal protease Kex2p. Computer analysis predicts a short coiled-coil domain in Luv1p. We propose that this protein maintains traffic through or the integrity of the early endosome and that this function is required for proper vacuolar morphology.

MeSH Terms
Amino Acid Sequence Calcineurin/metabolism Carrier Proteins/genetics,metabolism Cell Compartmentation Chemical Fractionation Cloning, Molecular Endosomes/metabolism Fungal Proteins/genetics,metabolism Genes, Fungal Golgi Apparatus/metabolism Green Fluorescent Proteins Luminescent Proteins/genetics,metabolism Molecular Sequence Data Mutagenesis Proprotein Convertases Saccharomyces cerevisiae/genetics,growth & development,metabolism,physiology Saccharomyces cerevisiae Proteins Subtilisins/metabolism Vacuoles/physiology Vesicular Transport Proteins
Chemicals
Carrier Proteins Fungal Proteins Luminescent Proteins Saccharomyces cerevisiae Proteins VPS54 protein, S cerevisiae Vesicular Transport Proteins Green Fluorescent Proteins Calcineurin Proprotein Convertases Subtilisins KEX2 protein, S cerevisiae
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Conboy M J
Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.
Cyert M S
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.11.7.2429
Published
2000-07-00
Pages
2429-43
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC14930
Subset
IM
Grants
NIGMS NIH HHS · R01 GM048729 · United States
NIGMS NIH HHS · T32 GM007276 · United States
NIGMS NIH HHS · 5 T32 GM07276 · United States
NIGMS NIH HHS · GM-48729 · United States
External Links
PubMed source DOI: 10.1091/mbc.11.7.2429 PMC Full Text View journal details
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