TY - JOUR TI - Intracellular functions and motile properties of bi-directional kinesin-5 Cin8 are regulated by neck linker docking AU - Goldstein-Levitin, Alina AU - Pandey, Himanshu AU - Allhuzaeel, Kanary AU - Kass, Itamar AU - Gheber, Larisa A2 - Verhey, Kristen J A2 - Akhmanova, Anna VL - 10 PY - 2021 DA - 2021/08/13 SP - e71036 C1 - eLife 2021;10:e71036 DO - 10.7554/eLife.71036 UR - https://doi.org/10.7554/eLife.71036 AB - 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. KW - kinesin-5 KW - mitotic spindle KW - microtubules KW - single molecule motility KW - antiparallel microtubule sliding KW - bi-directional motility JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -