Yeast dynamin Vps1 associates with clathrin to facilitate vesicular trafficking and controls Golgi homeostasis

Gadila, Shiva Kumar Goud; Williams, Michelle; Saimani, Uma; Cruz, Mariel Delgado; Makaraci, Pelin; Woodman, Sara; Short, John C. W.; McDermott, Hyoeun; Kim, Dong Kyu

doi:10.1016/j.ejcb.2017.02.004

Cited by 7 publications

(11 citation statements)

References 81 publications

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“…In addition, Vps1 has been reported to localize at endocytic sites on the plasma membrane, interact with endocytic proteins like clathrin, and influence the lifetimes and recruitment of endocytic proteins (Yu and Cai, 2004;Nannapaneni et al, 2010;Smaczynska-de Rooij et al, 2012). Others however, have failed to observe Vps1 at endocytic sites (Kishimoto et al, 2011;Goud Gadila et al, 2017), or observe a role in endocytic vesicle scission (Nothwehr et al, 1995;Kaksonen et al, 2005). The role of Vps1 in clathrin-mediated endocytosis thus remains debated.…”

Section: Introductionmentioning

confidence: 99%

Regulation of membrane scission in yeast endocytosis

Menon

Kaksonen

2021

Preprint

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During clathrin-mediated endocytosis, a flat plasma membrane is shaped into an invagination that undergoes scission to form a vesicle. In mammalian cells, the force that drives the transition from invagination to vesicle is primarily provided by the GTPase dynamin that acts in concert with crescent-shaped BAR domain proteins. In yeast cells, the mechanism of endocytic scission is unclear. The yeast BAR domain protein complex Rvs161/167 (Rvs) nevertheless plays an important role in this process: deletion of Rvs dramatically reduces scission efficiency. A mechanistic understanding of the influence of Rvs on scission however, remains incomplete. We used quantitative live-cell imaging and genetic manipulation to understand the recruitment and function of Rvs and other late-stage proteins at yeast endocytic sites. We found that arrival of Rvs at endocytic sites is timed by interaction of its BAR domain with specific membrane curvature. A second domain of Rvs167- the SH3 domain- affects localization efficiency of Rvs. We show that Myo3, one of the two type-I myosins in Saccharomyces cerevisiae, has a role in recruiting Rvs167 via the SH3 domain. Removal of the SH3 domain also affects assembly and disassembly of actin and impedes membrane invagination. Our results indicate that both BAR and SH3 domains are important for the role of Rvs as a regulator of scission. We tested other proteins implicated in vesicle formation in Saccharomyces cerevisiae , and found that neither synaptojanins nor dynamin contribute directly to membrane scission. We propose that recruitment of Rvs BAR domains delays scission and allows invaginations to grow by stabilizing them. We also propose that vesicle formation is dependent on the force exerted by the actin network component of the endocytic machinery.

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Section: Introductionmentioning

confidence: 99%

Regulation of membrane scission in yeast endocytosis

Menon

Kaksonen

2021

Preprint

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“…The three‐dimensional (3D) structure of a protein is crucial for its function, recruitment, and association with other proteins. Lines of evidence demonstrate that Vps1 locates at the TGN (Goud Gadila et al, ; Saimani et al, ), but its function there has been poorly understood. Therefore, finding its binding partners residing at the TGN can shed some light on how Vps1 may function at the TGN.…”

Section: Discussionmentioning

confidence: 99%

“…All plasmids used for this study are listed in Table . The DNA sequence of Vps1 or its fragments was fused to the GAL4 DNA‐binding domain (BD) of the bait plasmid pGKBT7 (KKD 0099), between the EcoR I and BamH I restriction sites (Goud Gadila et al, ) (Table ). DNA sequences that code for full‐length Ypt6, Ypt6 (1–72 amino acid), Ypt6 (73–142 amino acid), Ypt6 (143–215 amino acid), Vti1 (1–130 amino acid), Vti1 (1–186 amino acid), Vti1 (130–186 amino acid), Tlg1 (1–131 amino acid), Tlg2 (1–35 amino acid), Snc2 (1–52 amino acid), Snc2 (1–96 amino acid), Snc2 (28–52 amino acid), and Snc2 (52–96 amino acid) were inserted into the downstream of the GAL4 DNA‐activation domain (AD) (Stein et al, ) of the prey vector pGADT7 (KKD 0083) by using the EcoR I and BamH I restriction enzyme sites (Table ).…”

Section: Methodsmentioning

confidence: 99%

“…Goud Gadila et al (2017) pGBKT7-Vps1 (1-340 aa) KKD 0143 Obara et al (2013) pOK489 (mRFP-Cps1) KKD 0174…”

Section: Construction Of Plasmids and Yeast Strainsmentioning

confidence: 99%

“…Likewise, Vps1 was found to play an important role in homotypic vacuolar fusion (Peters et al, ) by inducing the interaction between Vam3 SNARE and the HOPS tethering complex at the vacuole (Kulkarni et al, ; Lurick et al, ). Furthermore, a recent study showing that loss of Vps1 leads to a robust increase of late Golgi in number indicates that Vps1 may function for homotypic fusion between late Golgi membranes (Goud Gadila et al, ). Identification and characterization of Vps1‐binding partners whose functions are implicated in tethering and fusion at the TGN could help understand the potential role of Vps1 for later steps of endosome‐to‐TGN trafficking.…”

Section: Introductionmentioning

confidence: 99%

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Yeast dynamin and Ypt6 function in parallel for the endosome‐to‐Golgi retrieval of Snc1

Makaraci

Cruz²,

McDermott

et al. 2019

Cell Biology International

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Protein recycling is an important cellular process required for cell homeostasis. Results from prior studies have shown that Vps1, a dynamin homologue in yeast, is implicated in protein recycling from the endosome to the trans-Golgi Network (TGN). However, the function of Vps1 in relation to Ypt6, a master GTPase in the recycling pathway, remains unknown. The present study reveals that Vps1 physically interacts with Ypt6 if at least one of them is full-length. We found that overexpression of full-length Vps1, but not GTP hydrolysis-defective Vps1 mutants, is sufficient to rescue abnormal phenotypes of Snc1 distribution provoked by loss of Ypt6, and vice versa. This suggests that Vps1 and Ypt6 function in parallel pathways instead of in a sequential pathway, and that GTP binding/hydrolysis of Vps1 is required for proper traffic of Snc1 toward the TGN. Additionally, we identified two novel Vps1 binding partners, Vti1 and Snc2, which function for the endosome-derived vesicle fusion at the TGN. Taken together, the present study demonstrates that Vps1 plays a role in later stages of the endosome-to-TGN traffic.

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