Interaction of the Saccharomyces cerevisiae Cortical Actin Patch Protein Rvs167p With Proteins Involved in ER to Golgi Vesicle Trafficking

Friesen, Helena; Colwill, Karen; Robertson, Karen; Schub, Oliver; Andrews, Brenda

doi:10.1534/genetics.104.040063

Cited by 22 publications

(35 citation statements)

References 75 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The SH3 domain of Rvs167p binds Gyp5p, whereas full-length Rvs161p appears to be required for its interaction with Gyp5p. The SH3 domain of Rvs167p binds Gyl1p and phosphorylation of Rvs167p inhibits this interaction (18,84,85,123,317,331). Rab GTPases cycle between an active GTP-bound form and an inactive GDP-bound form.…”

Section: Roles Of Rvs161p and Rvs167p In The Secretory Pathwaymentioning

confidence: 99%

The BAR Domain Proteins: Molding Membranes in Fission, Fusion, and Phagy

Ren

Vajjhala

Lee

et al. 2006

Microbiol Mol Biol Rev

192

186

View full text Add to dashboard Cite

SUMMARY The Bin1/amphiphysin/Rvs167 (BAR) domain proteins are a ubiquitous protein family. Genes encoding members of this family have not yet been found in the genomes of prokaryotes, but within eukaryotes, BAR domain proteins are found universally from unicellular eukaryotes such as yeast through to plants, insects, and vertebrates. BAR domain proteins share an N-terminal BAR domain with a high propensity to adopt α-helical structure and engage in coiled-coil interactions with other proteins. BAR domain proteins are implicated in processes as fundamental and diverse as fission of synaptic vesicles, cell polarity, endocytosis, regulation of the actin cytoskeleton, transcriptional repression, cell-cell fusion, signal transduction, apoptosis, secretory vesicle fusion, excitation-contraction coupling, learning and memory, tissue differentiation, ion flux across membranes, and tumor suppression. What has been lacking is a molecular understanding of the role of the BAR domain protein in each process. The three-dimensional structure of the BAR domain has now been determined and valuable insight has been gained in understanding the interactions of BAR domains with membranes. The cellular roles of BAR domain proteins, characterized over the past decade in cells as distinct as yeasts, neurons, and myocytes, can now be understood in terms of a fundamental molecular function of all BAR domain proteins: to sense membrane curvature, to bind GTPases, and to mold a diversity of cellular membranes.

show abstract

Section: Roles Of Rvs161p and Rvs167p In The Secretory Pathwaymentioning

confidence: 99%

The BAR Domain Proteins: Molding Membranes in Fission, Fusion, and Phagy

Ren

Vajjhala

Lee

et al. 2006

Microbiol Mol Biol Rev

192

186

View full text Add to dashboard Cite

show abstract

“…Affinity columns and Western blot analysis: Purified Abp1p SH3 domains were dialyzed into 20 mm HEPES (pH 8.0), 300 mm NaCl, and 10% glycerol and subsequently bound to AffiGel 10 resin (Bio-Rad, Hercules, CA) as previously described (Friesen et al 2005). The efficiency of the coupling reaction was determined by comparison of the supernatant before and after coupling, which was measured by Bradford assay and also visualized by 15% Tris-Tricine PAGE and subsequent Coomassie Blue staining of the polyacrylamide gel.…”

Section: By263 Abp1dtkanmentioning

confidence: 99%

“…The concentration of coupled SH3 domain on the resin was 1.4 mg/ml. Abp1p SH3 domain affinity columns were constructed as previously described (Friesen et al 2005).…”

Section: By263 Abp1dtkanmentioning

confidence: 99%

The Biologically Relevant Targets and Binding Affinity Requirements for the Function of the Yeast Actin-Binding Protein 1 Src-Homology 3 Domain Vary With Genetic Context

Haynes

Garcı́a²,

Stollar

et al. 2007

Genetics

View full text Add to dashboard Cite

Many protein-protein interaction domains bind to multiple targets. However, little is known about how the interactions of a single domain with many proteins are controlled and modulated under varying cellular conditions. In this study, we investigated the in vivo effects of Abp1p SH3 domain mutants that incrementally reduce target-binding affinity in four different yeast mutant backgrounds in which Abp1p activity is essential for growth. Although the severity of the phenotypic defects observed generally increased as binding affinity was reduced, some genetic backgrounds (prk1D and sla1D) tolerated large affinity reductions while others (sac6D and sla2D) were much more sensitive to these reductions. To elucidate the mechanisms behind these observations, we determined that Ark1p is the most important Abp1p SH3 domain interactor in prk1D cells, but that interactions with multiple targets, including Ark1p and Scp1p, are required in the sac6D background. We establish that the Abp1p SH3 domain makes different, functionally important interactions under different genetic conditions, and these changes in function are reflected by changes in the binding affinity requirement of the domain. These data provide the first evidence of biological relevance for any Abp1p SH3 domain-mediated interaction. We also find that considerable reductions in binding affinity are tolerated by the cell with little effect on growth rate, even when the actin cytoskeletal morphology is significantly perturbed.

show abstract

“…Equivalent amounts of each sample were analyzed by SDS-PAGE and transferred to nitrocellulose using a dry transfer apparatus. Coimmunoprecipitations were done as described previously (Friesen et al, 2005). Immunoblotting was done with antibodies against Rvs161p (gift from H. Riezman, University of Geneva, Geneva, Switzerland), Rvs167p and HA (12CA5).…”

Section: Protein Immunoprecipitation and Western Blottingmentioning

confidence: 99%

“…We have found that the rvs delete strains have synthetic growth defects in combination with two genes involved in vesicle trafficking from endoplasmic reticulum (ER) to Golgi, sec22⌬ and rud3⌬ (Newman and Ferro-Novick, 1990;Kim, 2003), suggesting that the Rvs proteins may be required for proper ER-to-Golgi trafficking. Additional evidence to support a role for Rvs167p in ER-to-Golgi vesicle trafficking comes from screens for proteins that interact with the SH3 domain of Rvs167p: the Rvs167p SH3 domain has been shown to bind to at least one protein involved in vesicle trafficking from ER to Golgi: Gyp5p (Bon et al, 2000;Ho et al, 2002;Chesneau et al, 2004;Friesen et al, 2005) as well as the Gyp5p-interacting protein Gyl1p (Chesneau et al, 2004;Friesen et al, 2005). In addition, the finding that RVS167 and RVS161 have synthetic genetic interactions with VPS21, a gene involved in the delivery of vacuolar and endocytosed proteins to the vacuole (Gerrard et al, 2000; interaction with RVS167 reported previously by SingerKrü ger and Ferro-Novick, 1997) suggests a role for Rvs161p-167p in multiple vesicle-mediated functions.…”

Section: Vesicle Traffickingmentioning

confidence: 99%

Characterization of the Yeast Amphiphysins Rvs161p and Rvs167p Reveals Roles for the Rvs Heterodimer In Vivo

Friesen

Humphries

et al. 2006

MBoC

Self Cite

View full text Add to dashboard Cite

We have used comprehensive synthetic lethal screens and biochemical assays to examine the biological role of the yeast amphiphysin homologues Rvs161p and Rvs167p, two proteins that play a role in regulation of the actin cytoskeleton, endocytosis, and sporulation. We found that unlike some forms of amphiphysin, Rvs161p-Rvs167p acts as an obligate heterodimer during vegetative growth and neither Rvs161p nor Rvs167p forms a homodimer in vivo. RVS161 and RVS167 have an identical set of 49 synthetic lethal interactions, revealing functions for the Rvs proteins in cell polarity, cell wall synthesis, and vesicle trafficking as well as a shared role in mating. Consistent with these roles, we show that the Rvs167p-Rvs161p heterodimer, like its amphiphysin homologues, can bind to phospholipid membranes in vitro, suggesting a role in vesicle formation and/or fusion. Our genetic screens also reveal that the interaction between Abp1p and the Rvs167p Src homology 3 (SH3) domain may be important under certain conditions, providing the first genetic evidence for a role for the SH3 domain of Rvs167p. Our studies implicate heterodimerization of amphiphysin family proteins in various functions related to cell polarity, cell integrity, and vesicle trafficking during vegetative growth and the mating response. INTRODUCTIONRVS161 and RVS167, which encode closely related proteins in Saccharomyces cerevisiae, were first identified in a screen for mutants that exhibited reduced viability upon starvation (Bauer et al., 1993). Mutation of RVS161 or RVS167 causes a phenotype consistent with a role for the Rvs proteins in cortical actin cytoskeleton organization and endocytosis: loss of viability and unusual cell morphology in poor growth medium or salt-containing medium, delocalized actin distribution under suboptimal growth conditions, abnormal (random) budding in diploids, and defects in endocytosis and sporulation (Bauer et al., 1993). Ultrastructural studies have revealed that rvs mutants accumulate late secretory vesicles at sites of membrane and cell wall construction (Breton et al., 2001), suggesting an additional role for Rvs161p and Rvs167p in vesicle trafficking.Rvs161p and Rvs167p are members of the BAR-domain family of proteins, which includes Bin1, Amphiphysin, and Rvs proteins (Sivadon et al., 1997). Amphiphysins are enriched in the mammalian brain and seem to function in synaptic vesicle endocytosis (for review, see Zhang and Zelhof, 2002). Bin1 is a splice isoform of amphiphysin 2 that has features of a tumor suppressor (Sakamuro et al., 1996). Proteins in this family are characterized by the presence of a conserved BAR domain, and it is through their BAR domains that Rvs161p interacts with Rvs167p (Navarro et al., 1997;Sivadon et al., 1997;Colwill et al., 1999). The crystal structure of the BAR domain of Drosophila amphiphysin has recently been solved (Peter et al., 2004). It is a crescentshaped dimer, in which each monomer forms a coiled coil. The curved shape is only revealed upon dimerization of the two slightly kinked ...

show abstract

Interaction of the Saccharomyces cerevisiae Cortical Actin Patch Protein Rvs167p With Proteins Involved in ER to Golgi Vesicle Trafficking

Cited by 22 publications

References 75 publications

The BAR Domain Proteins: Molding Membranes in Fission, Fusion, and Phagy

The BAR Domain Proteins: Molding Membranes in Fission, Fusion, and Phagy

The Biologically Relevant Targets and Binding Affinity Requirements for the Function of the Yeast Actin-Binding Protein 1 Src-Homology 3 Domain Vary With Genetic Context

Characterization of the Yeast Amphiphysins Rvs161p and Rvs167p Reveals Roles for the Rvs Heterodimer In Vivo

Contact Info

Product

Resources

About