Regulation and Possible Physiological Role of BI-Directional Motility of the Kinesin-5 Cin8

Ofer Shapira, Alina Goldstein, Jawdat Al-Bassam, Larisa Gheber

Research output: Contribution to journalMeeting Abstractpeer-review


The homoterameric bipolar kinesin-5 motors perform essential functions in
mitotic spindle dynamics by crosslinking and sliding apart antiparallel microtubules. S. cerevisiae cells express two kinesin-5s Cin8 and Kip1, which
overlap in function. We have recently demonstrated that Cin8 and Kip1 are
minus-end directed on the single-molecule level and can switch directionality
under a number of conditions (Duselder et al., 2015; Fridman et al., 2013;
Gerson-Gurwitz et al., 2011). The mechanism of this directionality switch
and its physiological significance remain unclear. We have also demonstrated
that Cin8 is differentially phosphorylated during late anaphase at three cyclindependent kinase 1 (Cdk1) sites located in its motor domain. This phosphorylation regulates Cin8 activity during anaphase (Avunie-Masala et al., 2011), but
its mechanism remains unclear.
Here we examined the in vitro motile properties and in vivo functions of
Cin8 by TIRF microscopy and live-cell imaging. We found that addition of
negative charge in a phospho-mimic Cin8 mutant weakens the MT-motor
interaction and regulates the motile properties and directionality of Cin8.
We also found that of the three Cdk1 sites in the catalytic domain of
Cin8, the S277 site contributes the most to regulation of Cin8 localization
and function during anaphase. Finally, we found that in vitro under high ionic
strength conditions, Cin8 not only moves to- but also clusters at the minus-end
of the MTs. This clustering causes Cin8 to reverse its directionality from fast
minus- to slow plus-end directed motility. Clustering of Cin8 at the minus-end
of the MTs serves as a primary site for capturing and antiparallel sliding of
MTs. Based on these results, we propose a revised model for activity of
Cin8 during mitosis and propose a physiological role for its minus-end
Original languageEnglish GB
Pages (from-to)460A-460A
JournalBiophysical Journal
Issue number3
StatePublished - 16 Feb 2016


Dive into the research topics of 'Regulation and Possible Physiological Role of BI-Directional Motility of the Kinesin-5 Cin8'. Together they form a unique fingerprint.

Cite this