title: Human kinetochore-associated kinesin CENP-E visualized at 17 A resolution bound to microtubules.
abstract: The highly dynamic process of cell division is effected, in part, by molecular motors that generate the forces necessary for its enactment. Several members of the kinesin superfamily of motor proteins are implicated in mitosis, such as CENP-E, which plays essential roles in cell division, including association with the kinetochore to stabilize attachment of chromosomes to microtubules prior to and during their separation. Neither the functional assembly state of CENP-E nor its direction of motion along the polar microtubule are certain. To determine the mode of interaction between CENP-E and microtubules, we have used cryo-electron microscopy to visualize CENP-E motor domains complexed with microtubules and calculated a density map of the complex to 17 A resolution by combining helical and single-particle reconstruction methods. The interface between the motor domain and microtubules was modeled by docking atomic-resolution models of the subunits into the cryoEM density map. Our results support a plus end motion for CENP-E, consistent with features of the crystallographic structure. Despite considerable functional differences from the monomeric transporter kinesin KIF1A and the oppositely directed ncd kinesin, CENP-E appears to share many features of the intermolecular interactions, suggesting that differences in motor function are governed by small variations in the loops at the microtubule interface.
authors.affiliation: Laboratoire de Microscopie Electronique Structurale, Institut de Biologie Structurale J.-P. Ebel, UMR 5075 CNRS-CEA-UJF, 38027 Grenoble, France.
authors.forename: E
authors.initials: E
authors.lastname: Neumann
authors.forename: I
authors.initials: I
authors.lastname: Garcia-Saez
authors.forename: S
authors.initials: S
authors.lastname: DeBonis
authors.forename: R H
authors.initials: RH
authors.lastname: Wade
authors.forename: F
authors.initials: F
authors.lastname: Kozielski
authors.forename: J F
authors.initials: JF
authors.lastname: Conway
mesh_terms: D000818:Animals; D002417:Cattle; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism; D020285:Cryoelectron Microscopy; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D021122:Protein Subunits; C068688:centromere protein E