Tether- scanning the kinesin motor domain reveals a core mechanical action

• 成果类型: Article
• 署名作者: Sumiyoshi, Rieko; Yamagishi, Masahiko; Furuta, Akane; Nishizaka, Takayuki; Furuta, Ken'ya; Cross, Robert A.; Yajima, Junichiro
• 署名单位: University of Tokyo; University of Tokyo; National Institute of Information & Communications Technology (NICT) - Japan; Gakushuin University; University of Warwick; University of Tokyo
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13243
• DOI: 10.1073/pnas.2403739121
• 发表日期: 2024-07-23
• 关键词: neck-linker; x-ray; conformational-changes; crystal-structure; microtubule; binding; direction; protein; ncd; processivity

摘要

Natural kinesin motors are tethered to their cargoes via short C- terminal or N- terminal linkers, whose docking against the core motor domain generates directional force. It remains unclear whether linker docking is the only process contributing directional force or whether linker docking is coupled to and amplifies an underlying, more fundamental force- generating mechanical cycle of the kinesin motor domain. Here, we show that kinesin motor domains tethered via double- stranded DNAs (dsDNAs) attached to surface loops drive robust microtubule (MT) gliding. Tethering using dsDNA attached to surface loops disconnects the C- terminal neck- linker and the effective attachment positions for the dsDNA tether are loop 2 or loop 10, which lie closest to the MT plus and minus ends, respectively. In three cases, we observed ancient, force- generating core mechanical action of the kinesin motor domain, which drives, and is amplified by, linker docking.