An interaction between Sla1p and Sla2p plays a role in regulating actin dynamics and endocytosis in budding yeast.
The importance of a dynamic actin cytoskeleton for facilitating endocytosis has been recognised for many years in budding yeast and is increasingly recognised in mammalian cells. However, the mechanism for actin recruitment and the role it plays in endocytosis is unclear. Here we show the importance of two yeast proteins in this process. We demonstrate that Sla1p and Sla2p interact in vitro and in vivo and that this interaction is mediated by the central domain of Sla2p, which includes its coiled-coil region, and by a domain of Sla1p between residues 118 and 361. Overexpression of the interacting fragment of Sla1p causes reduced fluid-phase endocytosis and, interestingly, defects in subsequent trafficking to vacuoles. We show that Sla2p is required for the polarised localisation of Sla1p in cells but not for its cortical localisation or for its overlapping localisation with actin. Generation of an Deltasla1Deltasla2 double mutant demonstrates that Sla2p is likely to act upstream of Sla1p in endocytosis, whereas sensitivity to latrunculin-A suggests that the proteins have opposite effects on actin dynamics. We propose that Sla2p recruits Sla1p to endocytic sites. Sla1p and its associated protein Pan1p then regulate actin assembly through interactions with Arp2/3 and Arp2/3-activating proteins Abp1p and Las17/Bee1p.
Institute of Biomedical and Life Sciences, Division of Biochemistry and Molecular Biology, Davidson Building, University of Glasgow, Scotland, UK.
Campbell W
CW
Gourlay
Hilary
H
Dewar
Derek T
DT
Warren
Rosaria
R
Costa
Nilima
N
Satish
Kathryn R
KR
Ayscough
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D051377:Actin-Related Protein 2; D000199:Actins / Q000378:metabolism*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D005656:Fungal Proteins / Q000378:metabolism; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D009154:Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure; D014617:Vacuoles / Q000378:metabolism; D051316:Wiskott-Aldrich Syndrome Protein
C062612:ABP1 protein, S cerevisiae; C477632:ARP2 protein, S cerevisiae; D051377:Actin-Related Protein 2; D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; C480361:LAS17 protein, S cerevisiae; D046911:Macromolecular Substances; D008840:Microfilament Proteins; C111870:PAN1 protein, S cerevisiae; C082265:SLA1 protein, S cerevisiae; C082266:SLA2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013844:Thiazoles; D053778:Thiazolidines; D051316:Wiskott-Aldrich Syndrome Protein; C037067:latrunculin A