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dc.contributor.authorDoxsey, Dylan D
dc.contributor.authorVeinotte, Kristen
dc.contributor.authorShen, Kuang
dc.date.accessioned2023-07-02T23:11:39Z
dc.date.available2023-07-02T23:11:39Z
dc.date.issued2022-11-03
dc.identifier.citationDoxsey DD, Veinotte K, Shen K. A New Crosslinking Assay to Study Guanine Nucleotide Binding in the Gtr Heterodimer of S. cerevisiae. Small GTPases. 2022 Jan;13(1):327-334. doi: 10.1080/21541248.2022.2141019. PMID: 36328771; PMCID: PMC9639563.en_US
dc.identifier.eissn2154-1256
dc.identifier.doi10.1080/21541248.2022.2141019en_US
dc.identifier.pmid36328771
dc.identifier.urihttp://hdl.handle.net/20.500.14038/52214
dc.description.abstractThe mechanistic target of rapamycin (mTOR) complex is responsible for coordinating nutrient availability with eukaryotic cell growth. Amino acid signals are transmitted towards mTOR via the Rag/Gtr heterodimers. Due to the obligatory heterodimeric architecture of the Rag/Gtr GTPases, investigating their biochemical properties has been challenging. Here, we describe an updated assay that allows us to probe the guanine nucleotide-binding affinity and kinetics to the Gtr heterodimers in Saccharomyces cerevisiae. We first identified the structural element that Gtr2p lacks to enable crosslinking. By using a sequence conservation-based mutation, we restored the crosslinking between Gtr2p and the bound nucleotides. Using this construct, we determined the nucleotide-binding affinities of the Gtr heterodimer, and found that it operates under a different form of intersubunit communication than human Rag GTPases. Our study defines the evolutionary divergence of the Gtr/Rag-mTOR axis of nutrient sensing.en_US
dc.language.isoenen_US
dc.relation.ispartofSmall GTPasesen_US
dc.relation.urlhttps://doi.org/10.1080/21541248.2022.2141019en_US
dc.rights© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.en_US
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectGTPaseen_US
dc.subjectGtr GTPaseen_US
dc.subjectRag GTPaseen_US
dc.subjectcrosslinkingen_US
dc.subjectenzyme kineticsen_US
dc.subjectguanine nucleotideen_US
dc.subjectmTORen_US
dc.subjectnutrient sensingen_US
dc.titleA New Crosslinking Assay to Study Guanine Nucleotide Binding in the Gtr Heterodimer of S. cerevisiaeen_US
dc.typeJournal Articleen_US
dc.source.journaltitleSmall GTPases
dc.source.volume13
dc.source.issue1
dc.source.beginpage327
dc.source.endpage334
dc.source.countryUnited States
dc.source.countryUnited States
dc.identifier.journalSmall GTPases
refterms.dateFOA2023-07-02T23:11:40Z
dc.contributor.departmentBiochemistry and Molecular Biotechnologyen_US
dc.contributor.departmentMorningside Graduate School of Biomedical Sciencesen_US
dc.contributor.departmentProgram in Molecular Medicineen_US
dc.contributor.studentDylan Doxsey


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© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
Except where otherwise noted, this item's license is described as © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.