Abstract
Exposure to mating pheromone in haploid Saccharomyces cerevisiae cells results in the arrest of the cell cycle, expression of mating-specific genes, and polarized growth toward the mating partner. Proteins involved in signaling, polarization, cell adhesion, and fusion are localized to the tip of the mating cell (shmoo) where fusion will eventually occur. The mechanisms ensuring the correct targeting and retention of these proteins are poorly understood. Here we show that in pheromone-treated cells, a reorganization of the plasma membrane involving lipid rafts results in the retention of proteins at the tip of the mating projection, segregated from the rest of the membrane. Sphingolipid and ergosterol biosynthetic mutants fail to polarize proteins to the tip of the shmoo and are deficient in mating. Our results show that membrane microdomain clustering at the mating projection is involved in the generation and maintenance of polarity during mating.
MeSH Terms
Acyltransferases/genetics,metabolism
Adaptor Proteins, Signal Transducing
Amino Acid Transport Systems
Carrier Proteins/genetics,metabolism
Cell Membrane/metabolism
Fungal Proteins/genetics,metabolism
Glucose Transport Proteins, Facilitative
Mating Factor
Membrane Microdomains/metabolism
Membrane Proteins/genetics,metabolism
Methyltransferases/genetics,metabolism
Monosaccharide Transport Proteins/genetics,metabolism
Peptides/metabolism
Phosphoric Monoester Hydrolases/genetics,metabolism
Recombinant Fusion Proteins/genetics,metabolism
Saccharomyces cerevisiae/metabolism,physiology
Saccharomyces cerevisiae Proteins/genetics,metabolism
Serine C-Palmitoyltransferase
Ubiquitin-Protein Ligases
ras GTPase-Activating Proteins/genetics,metabolism
Chemicals
Adaptor Proteins, Signal Transducing
Amino Acid Transport Systems
BUL1 protein, S cerevisiae
BUL2 protein, S cerevisiae
Carrier Proteins
FUS1 protein, S cerevisiae
Fungal Proteins
GAP1 protein, S cerevisiae
Glucose Transport Proteins, Facilitative
HXT2 protein, S cerevisiae
Membrane Proteins
Monosaccharide Transport Proteins
Peptides
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
ras GTPase-Activating Proteins
Mating Factor
Methyltransferases
delta 24-sterol methyltransferase
Acyltransferases
Serine C-Palmitoyltransferase
Ubiquitin-Protein Ligases
LCB3 protein, S cerevisiae
Phosphoric Monoester Hydrolases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bagnat Michel
Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
Simons Kai
References (39)
39 references, click to expand
-
Effect of sterol alterations on conjugation in Saccharomyces cerevisiae.
Yeast. 1992 Dec;8(12):1015-24
PMID: 1293881
-
The a-factor transporter (STE6 gene product) and cell polarity in the yeast Saccharomyces cerevisiae.
J Cell Biol. 1993 Mar;120(5):1203-15
PMID: 7679674
-
Fus2 localizes near the site of cell fusion and is required for both cell fusion and nuclear alignment during zygote formation.
J Cell Biol. 1995 Sep;130(6):1283-96
PMID: 7559752
-
Pheromone signalling and polarized morphogenesis in yeast.
Curr Opin Genet Dev. 1997 Feb;7(1):59-66
PMID: 9024634
-
Pheromone-regulated genes required for yeast mating differentiation.
J Cell Biol. 1998 Feb 9;140(3):461-83
PMID: 9456310
-
Rvs161p interacts with Fus2p to promote cell fusion in Saccharomyces cerevisiae.
J Cell Biol. 1998 May 4;141(3):567-84
PMID: 9566960
-
A role for the yeast actin cytoskeleton in pheromone receptor clustering and signalling.
Curr Biol. 1998 Jul 30-Aug 13;8(16):927-30
PMID: 9707405
-
New constructs and strategies for efficient PCR-based gene manipulations in yeast.
Yeast. 1998 Sep 15;14(12):1139-46
PMID: 9778799
-
Functions of lipid rafts in biological membranes.
Annu Rev Cell Dev Biol. 1998;14:111-36
PMID: 9891780
-
Cell polarity and morphogenesis in budding yeast.
Annu Rev Microbiol. 1998;52:687-744
PMID: 9891811
-
Components of a ubiquitin ligase complex specify polyubiquitination and intracellular trafficking of the general amino acid permease.
J Cell Biol. 2001 May 14;153(4):649-62
PMID: 11352928
-
Lipid rafts and signal transduction.
Nat Rev Mol Cell Biol. 2000 Oct;1(1):31-9
PMID: 11413487
-
Segregation of leading-edge and uropod components into specific lipid rafts during T cell polarization.
Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9642-7
PMID: 11493690
-
Cytoskeleton-dependent membrane domain segregation during neutrophil polarization.
Mol Biol Cell. 2001 Nov;12(11):3550-62
PMID: 11694588
-
The Ste5p scaffold.
J Cell Sci. 2001 Nov;114(Pt 22):3967-78
PMID: 11739629
-
Plasma membrane proton ATPase Pma1p requires raft association for surface delivery in yeast.
Mol Biol Cell. 2001 Dec;12(12):4129-38
PMID: 11739806
-
Septins: a ring to part mother and daughter.
Curr Genet. 2002 Jun;41(3):123-31
PMID: 12111093
-
The tight junction does not allow lipid molecules to diffuse from one epithelial cell to the next.
Nature. 1986 Aug 14-20;322(6080):639-41
PMID: 3748143
-
Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein.
Mol Cell Biol. 1987 Jul;7(7):2316-28
PMID: 3302672
-
The yeast gene ERG6 is required for normal membrane function but is not essential for biosynthesis of the cell-cycle-sparking sterol.
Mol Cell Biol. 1989 Aug;9(8):3447-56
PMID: 2677674
-
The yeast cell fusion protein FUS1 is O-glycosylated and spans the plasma membrane.
Proc Natl Acad Sci U S A. 1989 Dec;86(24):9916-20
PMID: 2690078
-
Courtship in Saccharomyces cerevisiae: an early cell-cell interaction during mating.
Mol Cell Biol. 1990 May;10(5):2202-13
PMID: 2183023
-
Courtship in S. cerevisiae: both cell types choose mating partners by responding to the strongest pheromone signal.
Cell. 1990 Nov 30;63(5):1039-51
PMID: 2257622
-
S. cerevisiae alpha pheromone receptors activate a novel signal transduction pathway for mating partner discrimination.
Cell. 1991 Oct 18;67(2):389-402
PMID: 1655282
-
A yeast gene (BEM1) necessary for cell polarization whose product contains two SH3 domains.
Nature. 1992 Mar 5;356(6364):77-9
PMID: 1538785
-
Membrane raft microdomains mediate front-rear polarity in migrating cells.
EMBO J. 1999 Nov 15;18(22):6211-20
PMID: 10562533
-
Specific sterols required for the internalization step of endocytosis in yeast.
Mol Biol Cell. 1999 Nov;10(11):3943-57
PMID: 10564282
-
Cell polarity in yeast.
Annu Rev Cell Dev Biol. 1999;15:365-91
PMID: 10611966
-
Lipid rafts function in biosynthetic delivery of proteins to the cell surface in yeast.
Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3254-9
PMID: 10716729
-
Sphingoid base synthesis requirement for endocytosis in Saccharomyces cerevisiae.
EMBO J. 2000 Jun 15;19(12):2824-33
PMID: 10856228
-
Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast.
Mol Cell. 2000 May;5(5):841-51
PMID: 10882120
-
G proteins mediate changes in cell shape by stabilizing the axis of polarity.
Mol Cell. 2000 May;5(5):853-64
PMID: 10882121
-
Yeast Cdc42 GTPase and Ste20 PAK-like kinase regulate Sho1-dependent activation of the Hog1 MAPK pathway.
EMBO J. 2000 Sep 1;19(17):4623-31
PMID: 10970855
-
Plasma membrane compartmentalization in yeast by messenger RNA transport and a septin diffusion barrier.
Science. 2000 Oct 13;290(5490):341-4
PMID: 11030653
-
A Saccharomyces gene family involved in invasive growth, cell-cell adhesion, and mating.
Proc Natl Acad Sci U S A. 2000 Oct 24;97(22):12158-63
PMID: 11027318
-
Prm1p, a pheromone-regulated multispanning membrane protein, facilitates plasma membrane fusion during yeast mating.
J Cell Biol. 2000 Oct 30;151(3):719-30
PMID: 11062271
-
Protein sorting upon exit from the endoplasmic reticulum.
Cell. 2001 Jan 26;104(2):313-20
PMID: 11207371
-
The immunological synapse.
Annu Rev Immunol. 2001;19:375-96
PMID: 11244041
-
Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression.
Nucleic Acids Res. 1994 Dec 25;22(25):5767-8
PMID: 7838736