Genetic and Biochemical Analyses of Yeast ESCRT

Banjade, Sudeep; Tang, Shaogeng; Emr, Scott D.

doi:10.1007/978-1-4939-9492-2_8

Cited by 8 publications

(12 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To test these hypotheses, we first utilized MVB cargo sorting assays (for cargoes Mup1 and Can1) in Saccharomyces cerevisiae ( Banjade et al, 2019a ; Banjade et al, 2019b ). In a vps24 ∆ strain overexpressing VPS2 , we observed that Mup1-pHluorin is sorted at about 40% compared to that of the wild type (WT), and that the canavanine sensitivity of vps24 ∆ is partially rescued ( Figure 1A ).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Design principles of the ESCRT-III Vps24-Vps2 module

Banjade

Shah

Tang

et al. 2021

eLife

Self Cite

View full text Add to dashboard Cite

ESCRT-III polymerization is required for all endosomal sorting complex required for transport (ESCRT)-dependent events in the cell. However, the relative contributions of the eight ESCRT-III subunits differ between each process. The minimal features of ESCRT-III proteins necessary for function and the role for the multiple ESCRT-III subunits remain unclear. To identify essential features of ESCRT-III subunits, we previously studied the polymerization mechanisms of two ESCRT-III subunits Snf7 and Vps24, identifying the association of the helix-4 region of Snf7 with the helix-1 region of Vps24 (Banjade et al., 2019a). Here, we find that mutations in the helix-1 region of another ESCRT-III subunit Vps2 can functionally replace Vps24 in Saccharomyces cerevisiae. Engineering and genetic selections revealed the required features of both subunits. Our data allow us to propose three minimal features required for ESCRT-III function – spiral formation, lateral association of the spirals through heteropolymerization, and binding to the AAA + ATPase Vps4 for dynamic remodeling.

show abstract

Section: Resultsmentioning

confidence: 99%

“…Lipid monolayers were prepared with a mixture of 60% POPC, 30% POPS, and 10% PI3P in chloroform. Carbon-coated electron microscope grids were used to make monolayers and incubate with proteins, as described before ( Banjade et al, 2019b ). Grids were stained with 2% ammonium molybdate and imaged on an FEI Morgagni 268 TEM.…”

Section: Methodsmentioning

confidence: 99%

Design principles of the ESCRT-III Vps24-Vps2 module

Banjade

Shah

Tang

et al. 2021

eLife

Self Cite

View full text Add to dashboard Cite

show abstract

“…To test these hypotheses, we first utilized MVB cargo sorting assays (for cargoes Mup1 and Can1) in S. cerevisiae (23,27). In a vps24∆ strain overexpressing VPS2, we observed that Mup1-pHluorin is sorted at about 40% compared to that of the wild type, and that the canavanine sensitivity of vps24∆ is partially rescued ( Figure 1A).…”

Section: Results: Overexpressing Vps2 Can Replace the Function Of Vpsmentioning

confidence: 99%

“…Lipid monolayers were prepared with a mixture of 60% POPC, 30% POPS and 10% PI3P in chloroform. Carbon-coated electron microscope grids were used to make monolayers and incubate with proteins, as described before (27). Grids were stained with 2% ammonium molybdate and imaged on an FEI Morgagni TEM.…”

Section: Electron Microscopymentioning

confidence: 99%

Design Principles of the ESCRT-III Vps24-Vps2 Module

Banjade

Shah

Tang³

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

ESCRT-III polymerization is required for all ESCRT-dependent events in the cell. However, the relative contributions of the eight ESCRT-III subunits differ between each process. The minimal features of ESCRT-III proteins necessary for function, and the role for the multiple ESCRT-III subunits remain unclear. To identify essential features of ESCRT-III subunits, we previously studied the polymerization mechanisms of two ESCRT-III subunits Snf7 and Vps24, identifying the association of the helix-4 region of Snf7 with the helix-1 region of Vps24 (Banjade et al., 2019). Here, we find that mutations in the helix-1 region of another ESCRT-III subunit Vps2 can functionally replace Vps24 in S. cerevisiae. Engineering and genetic selections revealed the required features of both subunits. Our data allow us to propose three minimal features required for ESCRT-III function – spiral formation, lateral association of the spirals through heteropolymerization, and binding to the AAA+ ATPase Vps4 for dynamic remodeling.

show abstract

“…ESCRTs are multiprotein complexes that recognize ubiquitinylated cargo as a sorting signal, and by directing them to the lysosome, allow the formation of intraluminal vesicles [100]. Four macromolecular complexes belong to ESCRT family: ESCRT0, ESCRTI, ESCRTII, and ESCRTIII, based on their appearance on the endosomal membrane [101]. ESCRT0 is composed of Vps27/HRS (HGF-regulated tyrosine kinase substrate) and Hse1/STAM (signal transducing adaptor molecule), which mediate interaction with both the endosomal PtdIns(3)P and ubiquitinated moieties of endocytosed receptors [102].…”

Section: Escrtsmentioning

confidence: 99%

Deliver on Time or Pay the Fine: Scheduling in Membrane Trafficking

Placidi

Campa

2021

IJMS

View full text Add to dashboard Cite

Membrane trafficking is all about time. Automation in such a biological process is crucial to ensure management and delivery of cellular cargoes with spatiotemporal precision. Shared molecular regulators and differential engagement of trafficking components improve robustness of molecular sorting. Sequential recruitment of low affinity protein complexes ensures directionality of the process and, concomitantly, serves as a kinetic proofreading mechanism to discriminate cargoes from the whole endocytosed material. This strategy helps cells to minimize losses and operating errors in membrane trafficking, thereby matching the appealed deadline. Here, we summarize the molecular pathways of molecular sorting, focusing on their timing and efficacy. We also highlight experimental procedures and genetic approaches to robustly probe these pathways, in order to guide mechanistic studies at the interface between biochemistry and quantitative biology.

show abstract

Genetic and Biochemical Analyses of Yeast ESCRT

Cited by 8 publications

References 15 publications

Design principles of the ESCRT-III Vps24-Vps2 module

Design principles of the ESCRT-III Vps24-Vps2 module

Design Principles of the ESCRT-III Vps24-Vps2 Module

Deliver on Time or Pay the Fine: Scheduling in Membrane Trafficking

Contact Info

Product

Resources

About