title: The MIS12 complex is a protein interaction hub for outer kinetochore assembly.
abstract: 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.
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology (IEO), Milan, Italy.
authors.forename: Arsen
authors.initials: A
authors.lastname: Petrovic
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Pasqualato
authors.forename: Prakash
authors.initials: P
authors.lastname: Dube
authors.forename: Veronica
authors.initials: V
authors.lastname: Krenn
authors.forename: Stefano
authors.initials: S
authors.lastname: Santaguida
authors.forename: Davide
authors.initials: D
authors.lastname: Cittaro
authors.forename: Silvia
authors.initials: S
authors.lastname: Monzani
authors.forename: Lucia
authors.initials: L
authors.lastname: Massimiliano
authors.forename: Jenny
authors.initials: J
authors.lastname: Keller
authors.forename: Aldo
authors.initials: A
authors.lastname: Tarricone
authors.forename: Alessio
authors.initials: A
authors.lastname: Maiolica
authors.forename: Holger
authors.initials: H
authors.lastname: Stark
authors.forename: Andrea
authors.initials: A
authors.lastname: Musacchio
mesh_terms: 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
chemical_list: 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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