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PMID:15651983

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Citation

Costa, R, Warren, DT and Ayscough, KR (2005) Lsb5p interacts with actin regulators Sla1p and Las17p, ubiquitin and Arf3p to couple actin dynamics to membrane trafficking processes. Biochem. J. 387:649-58

Abstract

The importance of coupling the process of endocytosis to factors that regulate actin dynamics has been clearly demonstrated in yeast, and many proteins involved in these mechanisms have been identified. Sla1p is a well-characterized yeast protein that binds both to activators of actin dynamics, Las17p and Pan1p, and to cargo proteins, such as the pheromone receptor Ste2p. Previously, we reported that the Lsb5 protein plays a role in endocytosis in yeast and that it localizes to the plasma membrane. Lsb5p has a similar structure to the GGA [Golgi-localized, gamma-ear-containing, Arf (ADP-ribosylation factor)-binding] family of proteins with an N-terminal VHS [Vps27p (vacuolar protein sorting protein 27), Hrs, Stam] domain and a GAT (GGA and Tom1) domain. It does not, however, contain either a gamma-adaptin ear or a clathrin-binding motif. In the present study, we have further defined its interaction site with both Sla1p and with Las17p, two regulators of actin dynamics. The site of interaction with Sla1p involves the Sla1 HD1 (homology domain 1), which also was shown previously to interact with the pheromone receptor Ste2p. We also demonstrate hitherto unknown interactions between Lsb5p and the active form of the yeast Arf3 protein, and with ubiquitin. Finally, we demonstrate a requirement for Arf3p expression in order to localize Lsb5p to the correct cortical site in cells. Taken together, our data provide further evidence for the role of Lsb5p in membrane-trafficking events at the plasma membrane and also demonstrate for the first time an interaction of Arf3 with the endocytic machinery in yeast.

Links

PubMed PMC1134994 Online version:10.1042/BJ20041729

Keywords

ADP-Ribosylation Factors/physiology; Actins/physiology; Carrier Proteins/physiology; Cell Membrane/metabolism; Cytoskeletal Proteins; Cytoskeleton/physiology; Endocytosis/physiology; Microfilament Proteins/physiology; Saccharomyces cerevisiae/physiology; Saccharomyces cerevisiae Proteins/physiology; Ubiquitin/physiology; Wiskott-Aldrich Syndrome Protein/physiology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:LSB5

located_in

GO:0030479: actin cortical patch

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:LSB5

located_in

GO:0005938: cell cortex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:LSB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P32790

F

Seeded From UniProt

complete

YEAST:LSB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P40994

F

Seeded From UniProt

complete

YEAST:LSB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q12446

F

Seeded From UniProt

complete

YEAST:SLA1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25369

F

Seeded From UniProt

complete

YEAST:ARF3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25369

F

Seeded From UniProt

complete

YEAST:LAS17

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25369

F

Seeded From UniProt

complete


See also

References

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