Noncanonical interaction with microtubules via the N-terminal nonmotor domain is critical for the functions of a bidirectional kinesin

Sudhir K. Singh, Nurit Siegler, Himanshu Pandey, Neta Yanir, Mary Popov, Alina Goldstein-Levitin, Mayan Sadan, Garrett Debs, Raz Zarivach, Gabriel A. Frank, Itamar Kass, Charles V. Sindelar, Ran Zalk, Larisa Gheber

Research output: Contribution to journalArticlepeer-review

Abstract

Several kinesin-5 motors (kinesin-5s) exhibit bidirectional motility. The mechanism of such motility remains unknown. Bidirectional kinesin-5s share a long N-terminal nonmotor domain (NTnmd), absent in exclusively plus-end–directed kinesins. Here, we combined in vivo, in vitro, and cryo–electron microscopy (cryo-EM) studies to examine the impact of NTnmd mutations on the motor functions of the bidirectional kinesin-5, Cin8. We found that NTnmd deletion mutants exhibited cell viability and spindle localization defects. Using cryo-EM, we examined the structure of a microtubule (MT)-bound motor domain of Cin8, containing part of its NTnmd. Modeling and molecular dynamic simulations based on the cryo-EM map suggested that the NTnmd of Cin8 interacts with the C-terminal tail of β-tubulin. In vitro experiments on subtilisin-treated MTs confirmed this notion. Last, we showed that NTnmd mutants are defective in plus-end–directed motility in single-molecule and antiparallel MT sliding assays. These findings demonstrate that the NTnmd, common to bidirectional kinesin-5s, is critical for their bidirectional motility and intracellular functions.

Original languageEnglish
Article numbereadi1367
JournalScience advances
Volume10
Issue number6
DOIs
StatePublished - 1 Feb 2024

ASJC Scopus subject areas

  • General

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