Intracellular functions and motile properties of bi-directional kinesin-5 Cin8 are regulated by neck linker docking

eLife

Kanary Allhuzaeel¹  Alina Goldstein-Levitin¹  Himanshu Pandey¹  Larisa Gheber¹  Itamar Kass² 

[1] Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel;Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, Israel;InterX LTD, Ramat-Gan, Israel;

关键词: kinesin-5;    mitotic spindle;    microtubules;    single molecule motility;    antiparallel microtubule sliding;    bi-directional motility;    S. cerevisiae;   

DOI  :  10.7554/eLife.71036

来源: eLife Sciences Publications, Ltd

【 摘 要 】

In this study, we analyzed intracellular functions and motile properties of neck-linker (NL) variants of the bi-directional S. cerevisiae kinesin-5 motor, Cin8. We also examined – by modeling – the configuration of H-bonds during NL docking. Decreasing the number of stabilizing H-bonds resulted in partially functional variants, as long as a conserved backbone H-bond at the N-latch position (proposed to stabilize the docked conformation of the NL) remained intact. Elimination of this conserved H-bond resulted in production of a non-functional Cin8 variant. Surprisingly, additional H-bond stabilization of the N-latch position, generated by replacement of the NL of Cin8 by sequences of the plus-end directed kinesin-5 Eg5, also produced a nonfunctional variant. In that variant, a single replacement of N-latch asparagine with glycine, as present in Cin8, eliminated the additional H-bond stabilization and rescued the functional defects. We conclude that exact N-latch stabilization during NL docking is critical for the function of bi-directional kinesin-5 Cin8.

【 授权许可】

CC BY