title: EB1 regulates attachment of Ska1 with microtubules by forming extended structures on the microtubule lattice.
abstract: Kinetochore couples chromosome movement to dynamic microtubules, a process that is fundamental to mitosis in all eukaryotes but poorly understood. In vertebrates, spindle-kinetochore-associated (Ska1-3) protein complex plays an important role in this process. However, the proteins that stabilize Ska-mediated kinetochore-microtubule attachment remain unknown. Here we show that microtubule plus-end tracking protein EB1 facilitates Ska localization on microtubules in vertebrate cells. EB1 depletion results in a significant reduction of Ska1 recruitment onto microtubules and defects in mitotic chromosome alignment, which is also reflected in computational modelling. Biochemical experiments reveal that EB1 interacts with Ska1, facilitates Ska1-microtubule attachment and together stabilizes microtubules. Structural studies reveal that EB1 either with Ska1 or Ska complex forms extended structures on microtubule lattice. Results indicate that EB1 promotes Ska association with K-fibres and facilitates kinetochore-microtubule attachment. They also implicate that in vertebrates, chromosome coupling to dynamic microtubules could be mediated through EB1-Ska extended structures.
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Geethu E
authors.initials: GE
authors.lastname: Thomas
authors.affiliation: School of Physics, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: K
authors.initials: K
authors.lastname: Bandopadhyay
authors.affiliation: Department of Solid State Physics, Indian Association for the Cultivation of Science, Kolkata 700032, India.
authors.forename: Sabyasachi
authors.initials: S
authors.lastname: Sutradhar
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: M R
authors.initials: MR
authors.lastname: Renjith
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Puja
authors.initials: P
authors.lastname: Singh
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: K K
authors.initials: KK
authors.lastname: Gireesh
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Steny
authors.initials: S
authors.lastname: Simon
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Binshad
authors.initials: B
authors.lastname: Badarudeen
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Hindol
authors.initials: H
authors.lastname: Gupta
authors.affiliation: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, New Delhi 110016, India.
authors.forename: Manidipa
authors.initials: M
authors.lastname: Banerjee
authors.affiliation: Department of Solid State Physics, Indian Association for the Cultivation of Science, Kolkata 700032, India.
authors.forename: Raja
authors.initials: R
authors.lastname: Paul
authors.affiliation: School of Physics, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: J
authors.identifier: 0000-0002-6633-9862
authors.initials: J
authors.lastname: Mitra
authors.affiliation: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus, Thiruvananthapuram 695016, India.
authors.forename: Tapas K
authors.initials: TK
authors.lastname: Manna
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D020090:Chromosome Segregation / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008938:Mitosis / Q000235:genetics; D034622:RNA Interference; D017386:Sequence Homology, Amino Acid
chemical_list: D002868:Chromosomal Proteins, Non-Histone; C500985:MAPRE1 protein, human; D008869:Microtubule-Associated Proteins; C516427:SKA1 protein, human