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

TY - JOUR

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

U2 - 10.1038/ncomms15822

DO - 10.1038/ncomms15822

M3 - Journal article

C2 - 28604727

SN - 2041-1723

VL - 8

JO - Nature Communications

JF - Nature Communications

M1 - 15822

ER -