title: Kinetochore-microtubule attachment throughout mitosis potentiated by the elongated stalk of the kinetochore kinesin CENP-E.
abstract: Centromere protein E (CENP-E) is a highly elongated kinesin that transports pole-proximal chromosomes during congression in prometaphase. During metaphase, it facilitates kinetochore-microtubule end-on attachment required to achieve and maintain chromosome alignment. In vitro CENP-E can walk processively along microtubule tracks and follow both growing and shrinking microtubule plus ends. Neither the CENP-E-dependent transport along microtubules nor its tip-tracking activity requires the unusually long coiled-coil stalk of CENP-E. The biological role for the CENP-E stalk has now been identified through creation of "Bonsai" CENP-E with significantly shortened stalk but wild-type motor and tail domains. We demonstrate that Bonsai CENP-E fails to bind microtubules in vitro unless a cargo is contemporaneously bound via its C-terminal tail. In contrast, both full-length and truncated CENP-E that has no stalk and tail exhibit robust motility with and without cargo binding, highlighting the importance of CENP-E stalk for its activity. Correspondingly, kinetochore attachment to microtubule ends is shown to be disrupted in cells whose CENP-E has a shortened stalk, thereby producing chromosome misalignment in metaphase and lagging chromosomes during anaphase. Together these findings establish an unexpected role of CENP-E elongated stalk in ensuring stability of kinetochore-microtubule attachments during chromosome congression and segregation.
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Vitre
authors.affiliation: Physiology Department, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104.
authors.forename: Nikita
authors.initials: N
authors.lastname: Gudimchuk
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Ranier
authors.initials: R
authors.lastname: Borda
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Cell Biology, Washington University in Saint Louis, St Louis, MO 63110WPI Institute for Cell and Material Sciences, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: John E
authors.initials: JE
authors.lastname: Heuser
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 dcleveland@ucsd.edu gekate@mail.med.upenn.edu.
authors.forename: Don W
authors.initials: DW
authors.lastname: Cleveland
authors.affiliation: Physiology Department, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 dcleveland@ucsd.edu gekate@mail.med.upenn.edu.
authors.forename: Ekaterina L
authors.initials: EL
authors.lastname: Grishchuk
mesh_terms: D000705:Anaphase; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000502:physiology*; D002877:Chromosomes, Human / Q000378:metabolism; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008677:Metaphase; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014982:Xenopus laevis
chemical_list: D002868:Chromosomal Proteins, Non-Histone; C068688:centromere protein E