ArticleEukaryotic cell2007
Split-ubiquitin two-hybrid assay to analyze protein-protein interactions at the endosome: application to Saccharomyces cerevisiae Bro1 interacting with ESCRT complexes, the Doa4 ubiquitin hydrolase, and the Rsp5 ubiquitin ligase.
Article in Eukaryotic cell, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 34 citations in OpenAlex.
- Bro1 family proteins harmonize cargo sorting with vesicle formation.BioEssays : news and reviews in molecular, cellular and developmental biology · 2022Article
- Bro1 stimulates Vps4 to promote intralumenal vesicle formation during multivesicular body biogenesis.The Journal of cell biology · 2021Article
- EhVps23: A Component of ESCRT-I That Participates in Vesicular Trafficking and Phagocytosis ofFrontiers in cellular and infection microbiology · 2021Article
- Arabidopsis ALIX is required for the endosomal localization of the deubiquitinating enzyme AMSH3.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Conserved Mode of Interaction between Yeast Bro1 Family V Domains and YP(X)nL Motif-Containing Target Proteins.Eukaryotic cell · 2015Article
- Host ESCRT proteins are required for bromovirus RNA replication compartment assembly and function.PLoS pathogens · 2015Article
- The ESCRT-III adaptor protein Bro1 controls functions of regulator for free ubiquitin chains 1 (Rfu1) in ubiquitin homeostasis.The Journal of biological chemistry · 2014Article
- The yeast Alix homolog Bro1 functions as a ubiquitin receptor for protein sorting into multivesicular endosomes.Developmental cell · 2013Article
- A dual role for K63-linked ubiquitin chains in multivesicular body biogenesis and cargo sorting.Molecular biology of the cell · 2012Article
- Alix protein is substrate of Ozz-E3 ligase and modulates actin remodeling in skeletal muscle.The Journal of biological chemistry · 2012Article
- The Phe105 loop of Alix Bro1 domain plays a key role in HIV-1 release.Structure (London, England : 1993) · 2011Article
- Proline-rich regions and motifs in trafficking: from ESCRT interaction to viral exploitation.Traffic (Copenhagen, Denmark) · 2011Review
- MVB vesicle formation: ESCRT-dependent, ESCRT-independent and everything in between.Current opinion in cell biology · 2011Review
- Chromatin modifying protein 1A (Chmp1A) of the endosomal sorting complex required for transport (ESCRT)-III family activates ataxia telangiectasia mutated (ATM) for PanC-1 cell growth inhibition.Cell cycle (Georgetown, Tex.) · 2011Article
- Intersection of the multivesicular body pathway and lipid homeostasis in RNA replication by a positive-strand RNA virus.Journal of virology · 2011Article
- A perturbed ubiquitin landscape distinguishes between ubiquitin in trafficking and in proteolysis.Molecular & cellular proteomics : MCP · 2011Article
- Protein-Protein Interaction Network and Subcellular Localization of the Arabidopsis Thaliana ESCRT Machinery.Frontiers in plant science · 2011Article
- Rescue of HIV-1 release by targeting widely divergent NEDD4-type ubiquitin ligases and isolated catalytic HECT domains to Gag.PLoS pathogens · 2010Article
- The ESCRT-associated protein Alix recruits the ubiquitin ligase Nedd4-1 to facilitate HIV-1 release through the LYPXnL L domain motif.Journal of virology · 2010Article
- Intervention of Bro1 in pH-responsive Rim20 localization in Saccharomyces cerevisiae.Eukaryotic cell · 2010Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Targeting of membrane proteins into the lysosomal/vacuolar lumen for degradation requires their prior sorting into multivesicular bodies (MVB). The MVB sorting pathway depends on ESCRT-0, -I, -II, and -III protein complexes functioning on the endosomal membrane and on additional factors, such as Bro1/Alix and the ubiquitin ligase Rsp5/Nedd4. We used the split-ubiquitin two-hybrid assay to analyze the interaction partners of yeast Bro1 at its natural cellular location. We show that Bro1 interacts with ESCRT-I and -III components, including Vps23, the Saccharomyces cerevisiae homologue of human Tsg101. These interactions do not require the C-terminal proline-rich domain (PRD) of Bro1. Rather, this PRD interacts with the Doa4 deubiquitinating enzyme to recruit it to the endosome. This interaction is disrupted by a single amino acid substitution in the conserved ELC box motif in Doa4. The PRD of Bro1 also mediates an association with Rsp5, and this interaction appears to be conserved, as Alix, the human homologue of Bro1, coimmunoprecipitates with Nedd4 in yeast lysates. We further show that the Bro1 PRD domain is essential to MVB sorting of only cargo proteins whose sorting to the vacuolar lumen is dependent on their own ubiquitination and Doa4. The Bro1 region preceding the PRD, however, is required for MVB sorting of proteins irrespective of whether their targeting to the vacuole is dependent on their ubiquitination and Doa4. Our data indicate that Bro1 interacts with several ESCRT components and contributes via its PRD to associating ubiquitinating and deubiquitinating enzymes with the MVB sorting machinery.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.