Structure of a Blinkin-BUBR1 complex reveals an interaction crucial for kinetochore-mitotic checkpoint regulation via an unanticipated binding Site

TY - JOUR

T1 - Structure of a Blinkin-BUBR1 complex reveals an interaction crucial for kinetochore-mitotic checkpoint regulation via an unanticipated binding Site

AU - Bolanos-Garcia, Victor M

AU - Lischetti, Tiziana

AU - Matak-Vinković, Dijana

AU - Cota, Ernesto

AU - Simpson, Pete J

AU - Chirgadze, Dimitri Y

AU - Spring, David R

AU - Robinson, Carol V

AU - Nilsson, Jakob

AU - Blundell, Tom L

N1 - Copyright © 2011 Elsevier Ltd. All rights reserved.

PY - 2011/11/9

Y1 - 2011/11/9

N2 - The maintenance of genomic stability relies on the spindle assembly checkpoint (SAC), which ensures accurate chromosome segregation by delaying the onset of anaphase until all chromosomes are properly bioriented and attached to the mitotic spindle. BUB1 and BUBR1 kinases are central for this process and by interacting with Blinkin, link the SAC with the kinetochore, the macromolecular assembly that connects microtubules with centromeric DNA. Here, we identify the Blinkin motif critical for interaction with BUBR1, define the stoichiometry and affinity of the interaction, and present a 2.2 resolution crystal structure of the complex. The structure defines an unanticipated BUBR1 region responsible for the interaction and reveals a novel Blinkin motif that undergoes a disorder-to-order transition upon ligand binding. We also show that substitution of several BUBR1 residues engaged in binding Blinkin leads to defects in the SAC, thus providing the first molecular details of the recognition mechanism underlying kinetochore-SAC signaling.

AB - The maintenance of genomic stability relies on the spindle assembly checkpoint (SAC), which ensures accurate chromosome segregation by delaying the onset of anaphase until all chromosomes are properly bioriented and attached to the mitotic spindle. BUB1 and BUBR1 kinases are central for this process and by interacting with Blinkin, link the SAC with the kinetochore, the macromolecular assembly that connects microtubules with centromeric DNA. Here, we identify the Blinkin motif critical for interaction with BUBR1, define the stoichiometry and affinity of the interaction, and present a 2.2 resolution crystal structure of the complex. The structure defines an unanticipated BUBR1 region responsible for the interaction and reveals a novel Blinkin motif that undergoes a disorder-to-order transition upon ligand binding. We also show that substitution of several BUBR1 residues engaged in binding Blinkin leads to defects in the SAC, thus providing the first molecular details of the recognition mechanism underlying kinetochore-SAC signaling.

KW - Amino Acid Sequence

KW - Binding Sites

KW - Cell Cycle Proteins

KW - Crystallography, X-Ray

KW - HeLa Cells

KW - Humans

KW - Kinetochores

KW - M Phase Cell Cycle Checkpoints

KW - Microtubule-Associated Proteins

KW - Mitosis

KW - Molecular Sequence Data

KW - Multiprotein Complexes

KW - Protein Binding

KW - Protein Interaction Domains and Motifs

KW - Protein-Serine-Threonine Kinases

KW - Two-Hybrid System Techniques

U2 - 10.1016/j.str.2011.09.017

DO - 10.1016/j.str.2011.09.017

M3 - Journal article

C2 - 22000412

SN - 0969-2126

VL - 19

SP - 1691

EP - 1700

JO - Structure

JF - Structure

IS - 11

ER -