The MIS12 complex is a protein interaction hub for outer kinetochore assembly.
Kinetochores are nucleoprotein assemblies responsible for the attachment of chromosomes to spindle microtubules during mitosis. The KMN network, a crucial constituent of the outer kinetochore, creates an interface that connects microtubules to centromeric chromatin. The NDC80, MIS12, and KNL1 complexes form the core of the KMN network. We recently reported the structural organization of the human NDC80 complex. In this study, we extend our analysis to the human MIS12 complex and show that it has an elongated structure with a long axis of approximately 22 nm. Through biochemical analysis, cross-linking-based methods, and negative-stain electron microscopy, we investigated the reciprocal organization of the subunits of the MIS12 complex and their contacts with the rest of the KMN network. A highlight of our findings is the identification of the NSL1 subunit as a scaffold supporting interactions of the MIS12 complex with the NDC80 and KNL1 complexes. Our analysis has important implications for understanding kinetochore organization in different organisms.
Petrovic
Arsen
A
Department of Experimental Oncology, European Institute of Oncology (IEO), Milan, Italy.
Pasqualato
Sebastiano
S
Dube
Prakash
P
Krenn
Veronica
V
Santaguida
Stefano
S
Cittaro
Davide
D
Monzani
Silvia
S
Massimiliano
Lucia
L
Keller
Jenny
J
Tarricone
Aldo
A
Maiolica
Alessio
A
Stark
Holger
H
Musacchio
Andrea
A
D000595:Amino Acid Sequence; D002875:Chromosomes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D008938:Mitosis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism
C472643:Knl1 protein, human; C492664:MIS12 protein, human; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins
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