Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56

Daniel Hayward; James Bancroft; Davinderpreet Mangat; Tatiana Alfonso-Pérez; Sholto Dugdale; Julia McCarthy; Francis A. Barr; Ulrike Gruneberg

doi:10.1083/JCB.201905026

Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56

Daniel Hayward, James Bancroft, Davinderpreet Mangat, Tatiana Alfonso-Pérez, Sholto Dugdale, Julia McCarthy, Francis A. Barr, Ulrike Gruneberg^*

^*Corresponding author for this work

Randall Centre of Cell & Molecular Biophysics

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

30 Citations (Scopus)

Abstract

During mitosis, the formation of microtubule-kinetochore attachments is monitored by the serine/threonine kinase monopolar spindle 1 (MPS1). MPS1 is recruited to unattached kinetochores where it phosphorylates KNL1, BUB1, and MAD1 to initiate the spindle assembly checkpoint. This arrests the cell cycle until all kinetochores have been stably captured by microtubules. MPS1 also contributes to the error correction process rectifying incorrect kinetochore attachments. MPS1 activity at kinetochores requires autophosphorylation at multiple sites including threonine 676 in the activation segment or "T-loop." We now demonstrate that the BUBR1-bound pool of PP2A-B56 regulates MPS1 T-loop autophosphorylation and hence activation status in mammalian cells. Overriding this regulation using phosphomimetic mutations in the MPS1 T-loop to generate a constitutively active kinase results in a prolonged mitotic arrest with continuous turnover of microtubule-kinetochore attachments. Dynamic regulation of MPS1 catalytic activity by kinetochore-localized PP2A-B56 is thus critical for controlled MPS1 activity and timely cell cycle progression.

Original language	English
Pages (from-to)	3188-3199
Number of pages	12
Journal	Journal of Cell Biology
Volume	218
Issue number	10
DOIs	https://doi.org/10.1083/JCB.201905026
Publication status	Published - 7 Oct 2019

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10.1083/JCB.201905026

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@article{a369fbe6cf654bc5b0793a41a44ffe40,

title = "Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56",

abstract = "During mitosis, the formation of microtubule-kinetochore attachments is monitored by the serine/threonine kinase monopolar spindle 1 (MPS1). MPS1 is recruited to unattached kinetochores where it phosphorylates KNL1, BUB1, and MAD1 to initiate the spindle assembly checkpoint. This arrests the cell cycle until all kinetochores have been stably captured by microtubules. MPS1 also contributes to the error correction process rectifying incorrect kinetochore attachments. MPS1 activity at kinetochores requires autophosphorylation at multiple sites including threonine 676 in the activation segment or {"}T-loop.{"} We now demonstrate that the BUBR1-bound pool of PP2A-B56 regulates MPS1 T-loop autophosphorylation and hence activation status in mammalian cells. Overriding this regulation using phosphomimetic mutations in the MPS1 T-loop to generate a constitutively active kinase results in a prolonged mitotic arrest with continuous turnover of microtubule-kinetochore attachments. Dynamic regulation of MPS1 catalytic activity by kinetochore-localized PP2A-B56 is thus critical for controlled MPS1 activity and timely cell cycle progression.",

author = "Daniel Hayward and James Bancroft and Davinderpreet Mangat and Tatiana Alfonso-P{\'e}rez and Sholto Dugdale and Julia McCarthy and Barr, {Francis A.} and Ulrike Gruneberg",

note = "Funding Information: We thank Stephen Taylor (University of Manchester, Manchester, UK) for Hela-Flp-In/TREx cells, Prasad Jallepalli (Memorial Sloan-Kettering Cancer Center, New York, NY) for SKA3 pS34 antibody, Jakob Nilsson (University of Copenhagen, Copenhagen, Denmark) for helpful advice regarding phosphomimetic mutants, Dr. Renaud Caous for HEC1-pSer55 antibody purification, and Iona Manley and Zo{\"e} Geraghty for critical reading of the manuscript. D. Hayward was supported by a Medical Research Council Senior Non-Clinical Research fellowship awarded to U. Gruneberg (MR/K006703/1) and T. Alfonso-P{\'e}rez by a Biotechnology and Biological Sciences Research Council Strategic LoLa grant (BB/M00354X/1). The authors declare no competing financial interests. Author contributions: Conceptualization: U. Gruneberg. Investigation: D. Hayward, J. Bancroft, T. Alfonso-P{\'e}rez, D. Mangat, S. Dugdale, and J. McCarthy. Funding acquisition: U. Gruneberg, F.A. Barr. Supervision: U. Gruneberg, F.A. Barr. Writing - original draft: U. Gruneberg. Writing - review and editing: T. Alfonso-P{\'e}rez, D. Hayward, J. Bancroft, F. A. Barr, U. Gruneberg. Funding Information: D. Hayward was supported by a Medical Research Council Senior Non-Clinical Research fellowship awarded to U. Grune-berg (MR/K006703/1) and T. Alfonso-P{\'e}rez by a Biotechnology and Biological Sciences Research Council Strategic LoLa grant (BB/M00354X/1). The authors declare no competing financial interests. Publisher Copyright: {\textcopyright} 2019 Hwang Fu et al.",

year = "2019",

month = oct,

day = "7",

doi = "10.1083/JCB.201905026",

language = "English",

volume = "218",

pages = "3188--3199",

journal = "Journal of Cell Biology",

issn = "0021-9525",

publisher = "Rockefeller University Press",

number = "10",

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TY - JOUR

T1 - Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56

AU - Hayward, Daniel

AU - Bancroft, James

AU - Mangat, Davinderpreet

AU - Alfonso-Pérez, Tatiana

AU - Dugdale, Sholto

AU - McCarthy, Julia

AU - Barr, Francis A.

AU - Gruneberg, Ulrike

N1 - Funding Information: We thank Stephen Taylor (University of Manchester, Manchester, UK) for Hela-Flp-In/TREx cells, Prasad Jallepalli (Memorial Sloan-Kettering Cancer Center, New York, NY) for SKA3 pS34 antibody, Jakob Nilsson (University of Copenhagen, Copenhagen, Denmark) for helpful advice regarding phosphomimetic mutants, Dr. Renaud Caous for HEC1-pSer55 antibody purification, and Iona Manley and Zoë Geraghty for critical reading of the manuscript. D. Hayward was supported by a Medical Research Council Senior Non-Clinical Research fellowship awarded to U. Gruneberg (MR/K006703/1) and T. Alfonso-Pérez by a Biotechnology and Biological Sciences Research Council Strategic LoLa grant (BB/M00354X/1). The authors declare no competing financial interests. Author contributions: Conceptualization: U. Gruneberg. Investigation: D. Hayward, J. Bancroft, T. Alfonso-Pérez, D. Mangat, S. Dugdale, and J. McCarthy. Funding acquisition: U. Gruneberg, F.A. Barr. Supervision: U. Gruneberg, F.A. Barr. Writing - original draft: U. Gruneberg. Writing - review and editing: T. Alfonso-Pérez, D. Hayward, J. Bancroft, F. A. Barr, U. Gruneberg. Funding Information: D. Hayward was supported by a Medical Research Council Senior Non-Clinical Research fellowship awarded to U. Grune-berg (MR/K006703/1) and T. Alfonso-Pérez by a Biotechnology and Biological Sciences Research Council Strategic LoLa grant (BB/M00354X/1). The authors declare no competing financial interests. Publisher Copyright: © 2019 Hwang Fu et al.

PY - 2019/10/7

Y1 - 2019/10/7

N2 - During mitosis, the formation of microtubule-kinetochore attachments is monitored by the serine/threonine kinase monopolar spindle 1 (MPS1). MPS1 is recruited to unattached kinetochores where it phosphorylates KNL1, BUB1, and MAD1 to initiate the spindle assembly checkpoint. This arrests the cell cycle until all kinetochores have been stably captured by microtubules. MPS1 also contributes to the error correction process rectifying incorrect kinetochore attachments. MPS1 activity at kinetochores requires autophosphorylation at multiple sites including threonine 676 in the activation segment or "T-loop." We now demonstrate that the BUBR1-bound pool of PP2A-B56 regulates MPS1 T-loop autophosphorylation and hence activation status in mammalian cells. Overriding this regulation using phosphomimetic mutations in the MPS1 T-loop to generate a constitutively active kinase results in a prolonged mitotic arrest with continuous turnover of microtubule-kinetochore attachments. Dynamic regulation of MPS1 catalytic activity by kinetochore-localized PP2A-B56 is thus critical for controlled MPS1 activity and timely cell cycle progression.

AB - During mitosis, the formation of microtubule-kinetochore attachments is monitored by the serine/threonine kinase monopolar spindle 1 (MPS1). MPS1 is recruited to unattached kinetochores where it phosphorylates KNL1, BUB1, and MAD1 to initiate the spindle assembly checkpoint. This arrests the cell cycle until all kinetochores have been stably captured by microtubules. MPS1 also contributes to the error correction process rectifying incorrect kinetochore attachments. MPS1 activity at kinetochores requires autophosphorylation at multiple sites including threonine 676 in the activation segment or "T-loop." We now demonstrate that the BUBR1-bound pool of PP2A-B56 regulates MPS1 T-loop autophosphorylation and hence activation status in mammalian cells. Overriding this regulation using phosphomimetic mutations in the MPS1 T-loop to generate a constitutively active kinase results in a prolonged mitotic arrest with continuous turnover of microtubule-kinetochore attachments. Dynamic regulation of MPS1 catalytic activity by kinetochore-localized PP2A-B56 is thus critical for controlled MPS1 activity and timely cell cycle progression.

UR - http://www.scopus.com/inward/record.url?scp=85072993970&partnerID=8YFLogxK

U2 - 10.1083/JCB.201905026

DO - 10.1083/JCB.201905026

M3 - Article

C2 - 31511308

AN - SCOPUS:85072993970

SN - 0021-9525

VL - 218

SP - 3188

EP - 3199

JO - Journal of Cell Biology

JF - Journal of Cell Biology

IS - 10

ER -

Checkpoint signaling and error correction require regulation of the MPS1 T-loop by PP2A-B56

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