Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling

Gang Zhang; Thomas Kruse; Blanca López-Méndez; Kathrine Beck Sylvestersen; Dimitriya H Garvanska; Simone Schopper; Michael Lund Nielsen; Jakob Nilsson

doi:10.1038/ncomms15822

Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling

Gang Zhang, Thomas Kruse, Blanca López-Méndez, Kathrine Beck Sylvestersen, Dimitriya H Garvanska, Simone Schopper, Michael Lund Nielsen, Jakob Nilsson

Novo Nordisk Foundation Center for Protein Research

42 Citations (Scopus)

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Abstract

Proper segregation of chromosomes depends on a functional spindle assembly checkpoint (SAC) and requires kinetochore localization of the Bub1 and Mad1/Mad2 checkpoint proteins. Several aspects of Mad1/Mad2 kinetochore recruitment in human cells are unclear and in particular the underlying direct interactions. Here we show that conserved domain 1 (CD1) in human Bub1 binds directly to Mad1 and a phosphorylation site exists in CD1 that stimulates Mad1 binding and SAC signalling. Importantly, fusion of minimal kinetochore-targeting Bub1 fragments to Mad1 bypasses the need for CD1, revealing that the main function of Bub1 is to position Mad1 close to KNL1 MELT repeats. Furthermore, we identify residues in Mad1 that are critical for Mad1 functionality, but not Bub1 binding, arguing for a direct role of Mad1 in the checkpoint. This work dissects functionally relevant molecular interactions required for spindle assembly checkpoint signalling at kinetochores in human cells.

Original language	English
Article number	15822
Journal	Nature Communications
Volume	8
Number of pages	12
ISSN	2041-1723
DOIs	https://doi.org/10.1038/ncomms15822
Publication status	Published - 12 Jun 2017

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Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signallingFinal published version, 1.99 MBLicence: CC BY

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title = "Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling",

abstract = "Proper segregation of chromosomes depends on a functional spindle assembly checkpoint (SAC) and requires kinetochore localization of the Bub1 and Mad1/Mad2 checkpoint proteins. Several aspects of Mad1/Mad2 kinetochore recruitment in human cells are unclear and in particular the underlying direct interactions. Here we show that conserved domain 1 (CD1) in human Bub1 binds directly to Mad1 and a phosphorylation site exists in CD1 that stimulates Mad1 binding and SAC signalling. Importantly, fusion of minimal kinetochore-targeting Bub1 fragments to Mad1 bypasses the need for CD1, revealing that the main function of Bub1 is to position Mad1 close to KNL1 MELT repeats. Furthermore, we identify residues in Mad1 that are critical for Mad1 functionality, but not Bub1 binding, arguing for a direct role of Mad1 in the checkpoint. This work dissects functionally relevant molecular interactions required for spindle assembly checkpoint signalling at kinetochores in human cells.",

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T1 - Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling

AU - Zhang, Gang

AU - Kruse, Thomas

AU - López-Méndez, Blanca

AU - Sylvestersen, Kathrine Beck

AU - Garvanska, Dimitriya H

AU - Schopper, Simone

AU - Nielsen, Michael Lund

AU - Nilsson, Jakob

PY - 2017/6/12

Y1 - 2017/6/12

N2 - Proper segregation of chromosomes depends on a functional spindle assembly checkpoint (SAC) and requires kinetochore localization of the Bub1 and Mad1/Mad2 checkpoint proteins. Several aspects of Mad1/Mad2 kinetochore recruitment in human cells are unclear and in particular the underlying direct interactions. Here we show that conserved domain 1 (CD1) in human Bub1 binds directly to Mad1 and a phosphorylation site exists in CD1 that stimulates Mad1 binding and SAC signalling. Importantly, fusion of minimal kinetochore-targeting Bub1 fragments to Mad1 bypasses the need for CD1, revealing that the main function of Bub1 is to position Mad1 close to KNL1 MELT repeats. Furthermore, we identify residues in Mad1 that are critical for Mad1 functionality, but not Bub1 binding, arguing for a direct role of Mad1 in the checkpoint. This work dissects functionally relevant molecular interactions required for spindle assembly checkpoint signalling at kinetochores in human cells.

AB - Proper segregation of chromosomes depends on a functional spindle assembly checkpoint (SAC) and requires kinetochore localization of the Bub1 and Mad1/Mad2 checkpoint proteins. Several aspects of Mad1/Mad2 kinetochore recruitment in human cells are unclear and in particular the underlying direct interactions. Here we show that conserved domain 1 (CD1) in human Bub1 binds directly to Mad1 and a phosphorylation site exists in CD1 that stimulates Mad1 binding and SAC signalling. Importantly, fusion of minimal kinetochore-targeting Bub1 fragments to Mad1 bypasses the need for CD1, revealing that the main function of Bub1 is to position Mad1 close to KNL1 MELT repeats. Furthermore, we identify residues in Mad1 that are critical for Mad1 functionality, but not Bub1 binding, arguing for a direct role of Mad1 in the checkpoint. This work dissects functionally relevant molecular interactions required for spindle assembly checkpoint signalling at kinetochores in human cells.

KW - Journal Article

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DO - 10.1038/ncomms15822

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VL - 8

JO - Nature Communications

JF - Nature Communications

M1 - 15822

ER -

Bub1 positions Mad1 close to KNL1 MELT repeats to promote checkpoint signalling

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