Epsin N-terminal homology domains perform an essential function regulating Cdc42 through binding Cdc42 GTPase-activating proteins

Rubén C Aguilar; Silvia A Longhi; Jonathan D Shaw; Lan-Yu Yeh; Sean Kim; Arne Schön; Ernesto Freire; Ariel Hsu; William K McCormick; Hadiya A Watson; Beverly Wendland

doi:10.1073/pnas.0510513103

Epsin N-terminal homology domains perform an essential function regulating Cdc42 through binding Cdc42 GTPase-activating proteins

Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4116-21. doi: 10.1073/pnas.0510513103. Epub 2006 Mar 6.

Authors

Rubén C Aguilar¹, Silvia A Longhi, Jonathan D Shaw, Lan-Yu Yeh, Sean Kim, Arne Schön, Ernesto Freire, Ariel Hsu, William K McCormick, Hadiya A Watson, Beverly Wendland

Affiliation

¹ Department of Biology, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.

Abstract

Epsins are endocytic proteins with a structured epsin N-terminal homology (ENTH) domain that binds phosphoinositides and a poorly structured C-terminal region that interacts with ubiquitin and endocytic machinery, including clathrin and endocytic scaffolding proteins. Yeast has two redundant genes encoding epsins, ENT1 and ENT2; deleting both genes is lethal. We demonstrate that the ENTH domain is both necessary and sufficient for viability of ent1Deltaent2Delta cells. Mutational analysis of the ENTH domain revealed a surface patch that is essential for viability and that binds guanine nucleotide triphosphatase-activating proteins for Cdc42, a critical regulator of cell polarity in all eukaryotes. Furthermore, the epsins contribute to regulation of specific Cdc42 signaling pathways in yeast cells. These data support a model in which the epsins function as spatial and temporal coordinators of endocytosis and cell polarity.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Adaptor Proteins, Signal Transducing / chemistry*
Adaptor Proteins, Signal Transducing / genetics
Adaptor Proteins, Signal Transducing / metabolism*
Adaptor Proteins, Vesicular Transport
Carrier Proteins / chemistry*
Carrier Proteins / genetics
Carrier Proteins / metabolism*
Cell Polarity
Endocytosis
Genes, Fungal
Models, Molecular
Mutation
Phenotype
Protein Structure, Tertiary
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins / chemistry*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism*
Vesicular Transport Proteins
cdc42 GTP-Binding Protein, Saccharomyces cerevisiae / metabolism*

Substances

Adaptor Proteins, Signal Transducing
Adaptor Proteins, Vesicular Transport
Carrier Proteins
ENT1 protein, S cerevisiae
ENT2 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
Vesicular Transport Proteins
cdc42 GTP-Binding Protein, Saccharomyces cerevisiae

Grants and funding

R01 GM-60979/GM/NIGMS NIH HHS/United States