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PMID: 10564282 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specific sterols required for the internalization step of endocytosis in yeast.

Molecular biology of the cell ·Vol. 10 ·No. 11 ·1999-11-00 ·Pages 3943-57

Munn AL, Heese-Peck A, Stevenson BJ, Pichler H, Riezman H

Abstract

Sterols are major components of the plasma membrane, but their functions in this membrane are not well understood. We isolated a mutant defective in the internalization step of endocytosis in a gene (ERG2) encoding a C-8 sterol isomerase that acts in the late part of the ergosterol biosynthetic pathway. In the absence of Erg2p, yeast cells accumulate sterols structurally different from ergosterol, which is the major sterol in wild-type yeast. To investigate the structural requirements of ergosterol for endocytosis in more detail, several erg mutants (erg2Delta, erg6Delta, and erg2Deltaerg6Delta) were made. Analysis of fluid phase and receptor-mediated endocytosis indicates that changes in the sterol composition lead to a defect in the internalization step. Vesicle formation and fusion along the secretory pathway were not strongly affected in the ergDelta mutants. The severity of the endocytic defect correlates with changes in sterol structure and with the abundance of specific sterols in the ergDelta mutants. Desaturation of the B ring of the sterol molecules is important for the internalization step. A single desaturation at C-8,9 was not sufficient to support internalization at 37 degrees C whereas two double bonds, either at C-5,6 and C-7,8 or at C-5,6 and C-8,9, allowed internalization.

MeSH Terms
Carboxypeptidases/metabolism Cathepsin A Cloning, Molecular Endocytosis/genetics Ergosterol/biosynthesis Glycoside Hydrolases/metabolism Kinetics Mating Factor Microscopy, Phase-Contrast Molecular Structure Mutation Peptides/metabolism Steroid Isomerases/genetics Sterols/chemistry,metabolism Yeasts/genetics beta-Fructofuranosidase
Chemicals
Peptides Sterols Mating Factor Glycoside Hydrolases beta-Fructofuranosidase Carboxypeptidases Cathepsin A Steroid Isomerases delta(8)-delta(7)-sterol isomerase Ergosterol
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Munn A L
Biozentrum of the University of Basel, CH-4056 Basel, Switzerland.
Heese-Peck A
Stevenson B J
Pichler H
Riezman H
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.10.11.3943
Published
1999-11-00
Pages
3943-57
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25690
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.10.11.3943 PMC Full Text View journal details
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