EB1 regulates attachment of Ska1 with microtubules by forming extended structures on the microtubule lattice

Geethu E. Thomas, K. Bandopadhyay, Sabyasachi Sutradhar, M. R. Renjith, Puja Singh, K. K. Gireesh, Steny Simon, Binshad Badarudeen, Hindol Gupta, Manidipa Banerjee, Raja Paul, J. Mitra and Tapas K. Manna ()
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Geethu E. Thomas: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
K. Bandopadhyay: School of Physics, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Sabyasachi Sutradhar: Indian Association for the Cultivation of Science
M. R. Renjith: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Puja Singh: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
K. K. Gireesh: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Steny Simon: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Binshad Badarudeen: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Hindol Gupta: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Manidipa Banerjee: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi
Raja Paul: Indian Association for the Cultivation of Science
J. Mitra: School of Physics, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus
Tapas K. Manna: School of Biology, Indian Institute of Science Education and Research Thiruvananthapuram, CET Campus

Nature Communications, 2016, vol. 7, issue 1, 1-13

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11665

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