TOR complex 1 negatively regulates NDR kinase Cbk1 to control cell separation in budding yeast

Magdalena Foltman; Iván Mendez; Joan J. Bech-Serra; Carolina De La Torre; Jennifer L. Brace; Eric L. Weiss; María Lucas; Ethel Queralt; Alberto Sanchez-Diaz

doi:10.1371/journal.pbio.3002263

TOR complex 1 negatively regulates NDR kinase Cbk1 to control cell separation in budding yeast

Magdalena Foltman^*, Iván Mendez, Joan J. Bech-Serra, Carolina De La Torre, Jennifer L. Brace, Eric L. Weiss, María Lucas, Ethel Queralt, Alberto Sanchez-Diaz^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

The target of rapamycin (TOR) signalling pathway plays a key role in the coordination between cellular growth and the cell cycle machinery in eukaryotes. The underlying molecular mechanisms by which TOR might regulate events after anaphase remain unknown. We show for the first time that one of the 2 TOR complexes in budding yeast, TORC1, blocks the separation of cells following cytokinesis by phosphorylation of a member of the NDR (nuclear Dbf2-related) protein-kinase family, the protein Cbk1. We observe that TORC1 alters the phosphorylation pattern of Cbk1 and we identify a residue within Cbk1 activation loop, T574, for which a phosphomimetic substitution makes Cbk1 catalytically inactive and indeed, reproduces TORC1 control over cell separation. In addition, we identify the exocyst component Sec3 as a key substrate of Cbk1, since Sec3 activates the SNARE complex to promote membrane fusion. TORC1 activity ultimately compromises the interaction between Sec3 and a t-SNARE component. Our data indicate that TORC1 negatively regulates cell separation in budding yeast by participating in Cbk1 phosphorylation, which in turn controls the fusion of secretory vesicles transporting hydrolase at the site of division.

Original language	English (US)
Article number	e3002263
Journal	PLoS biology
Volume	21
Issue number	8 August
DOIs	https://doi.org/10.1371/journal.pbio.3002263
State	Published - Aug 2023

Funding

ASD acknowledges a grant from the Consejerı ´a de Universidades, Investigacio ´ n, Medio Ambiente y Polı ´ tica Social del Gobierno de Cantabria, and anothergrantfromSociedadparaelDesarrollode Cantabria(SODERCAN).EQwasfundedby GeneralitatValenciana(CIDEGENT2020/41).ML acknowledgesresearchsupportbygrantRTI2018-097801-B-I00fundedbyMCIN/AEI/10.13039/ 501100011033andby“ERDFAwayofmaking Europe”andgrantPID2021-122611NB-100 fundedbyMCIN/AEI/10.13039/501100011033 andby“ESFInvestinginyourfuture”.Thefunders hadnoroleinstudydesign,datacollectionand analysis,decisiontopublish,orpreparationofthe manuscript. Funding: The Agencia Estatal de Investigacion (AEI) of Ministerio de Ciencia e Innovacion (MCIN) funded this work. Grants PID2019-106745GB-I00 funded by MCIN/AEI/10.13039/501100011033 to ASD, PID2019-109027GB-I00 funded by MCIN/AEI/10.13039/501100011033 to EQ. Moreover ASD acknowledges a grant from the Consejeria de Universidades, Investigacion, Medio Ambiente y Politica Social del Gobierno de Cantabria, and another grant from Sociedad para el Desarrollo de Cantabria (SODERCAN). EQ was funded by Generalitat Valenciana (CIDEGENT2020/41). ML acknowledges research support by grant RTI2018-097801-B-I00 funded by MCIN/AEI/ 10.13039/501100011033 and by ERDF A way of making Europe and grant PID2021-122611NB-100 funded by MCIN/AEI/ 10.13039/501100011033 and by ESF Investing in your future

ASJC Scopus subject areas

General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology
General Neuroscience
General Agricultural and Biological Sciences

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title = "TOR complex 1 negatively regulates NDR kinase Cbk1 to control cell separation in budding yeast",

abstract = "The target of rapamycin (TOR) signalling pathway plays a key role in the coordination between cellular growth and the cell cycle machinery in eukaryotes. The underlying molecular mechanisms by which TOR might regulate events after anaphase remain unknown. We show for the first time that one of the 2 TOR complexes in budding yeast, TORC1, blocks the separation of cells following cytokinesis by phosphorylation of a member of the NDR (nuclear Dbf2-related) protein-kinase family, the protein Cbk1. We observe that TORC1 alters the phosphorylation pattern of Cbk1 and we identify a residue within Cbk1 activation loop, T574, for which a phosphomimetic substitution makes Cbk1 catalytically inactive and indeed, reproduces TORC1 control over cell separation. In addition, we identify the exocyst component Sec3 as a key substrate of Cbk1, since Sec3 activates the SNARE complex to promote membrane fusion. TORC1 activity ultimately compromises the interaction between Sec3 and a t-SNARE component. Our data indicate that TORC1 negatively regulates cell separation in budding yeast by participating in Cbk1 phosphorylation, which in turn controls the fusion of secretory vesicles transporting hydrolase at the site of division.",

author = "Magdalena Foltman and Iv{\'a}n Mendez and Bech-Serra, {Joan J.} and {De La Torre}, Carolina and Brace, {Jennifer L.} and Weiss, {Eric L.} and Mar{\'i}a Lucas and Ethel Queralt and Alberto Sanchez-Diaz",

note = "Publisher Copyright: {\textcopyright} 2023 Foltman et al.",

year = "2023",

month = aug,

doi = "10.1371/journal.pbio.3002263",

language = "English (US)",

volume = "21",

journal = "PLoS biology",

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T1 - TOR complex 1 negatively regulates NDR kinase Cbk1 to control cell separation in budding yeast

AU - Foltman, Magdalena

AU - Mendez, Iván

AU - Bech-Serra, Joan J.

AU - De La Torre, Carolina

AU - Brace, Jennifer L.

AU - Weiss, Eric L.

AU - Lucas, María

AU - Queralt, Ethel

AU - Sanchez-Diaz, Alberto

PY - 2023/8

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N2 - The target of rapamycin (TOR) signalling pathway plays a key role in the coordination between cellular growth and the cell cycle machinery in eukaryotes. The underlying molecular mechanisms by which TOR might regulate events after anaphase remain unknown. We show for the first time that one of the 2 TOR complexes in budding yeast, TORC1, blocks the separation of cells following cytokinesis by phosphorylation of a member of the NDR (nuclear Dbf2-related) protein-kinase family, the protein Cbk1. We observe that TORC1 alters the phosphorylation pattern of Cbk1 and we identify a residue within Cbk1 activation loop, T574, for which a phosphomimetic substitution makes Cbk1 catalytically inactive and indeed, reproduces TORC1 control over cell separation. In addition, we identify the exocyst component Sec3 as a key substrate of Cbk1, since Sec3 activates the SNARE complex to promote membrane fusion. TORC1 activity ultimately compromises the interaction between Sec3 and a t-SNARE component. Our data indicate that TORC1 negatively regulates cell separation in budding yeast by participating in Cbk1 phosphorylation, which in turn controls the fusion of secretory vesicles transporting hydrolase at the site of division.

AB - The target of rapamycin (TOR) signalling pathway plays a key role in the coordination between cellular growth and the cell cycle machinery in eukaryotes. The underlying molecular mechanisms by which TOR might regulate events after anaphase remain unknown. We show for the first time that one of the 2 TOR complexes in budding yeast, TORC1, blocks the separation of cells following cytokinesis by phosphorylation of a member of the NDR (nuclear Dbf2-related) protein-kinase family, the protein Cbk1. We observe that TORC1 alters the phosphorylation pattern of Cbk1 and we identify a residue within Cbk1 activation loop, T574, for which a phosphomimetic substitution makes Cbk1 catalytically inactive and indeed, reproduces TORC1 control over cell separation. In addition, we identify the exocyst component Sec3 as a key substrate of Cbk1, since Sec3 activates the SNARE complex to promote membrane fusion. TORC1 activity ultimately compromises the interaction between Sec3 and a t-SNARE component. Our data indicate that TORC1 negatively regulates cell separation in budding yeast by participating in Cbk1 phosphorylation, which in turn controls the fusion of secretory vesicles transporting hydrolase at the site of division.

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TOR complex 1 negatively regulates NDR kinase Cbk1 to control cell separation in budding yeast

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