Examining kinesin processivity within a general gating framework

Johan O.L. Andreasson, Bojan Milic, Geng Yuan Chen, Nicholas R. Guydosh, William O. Hancock, Steven M. Block

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75 Scopus citations

Abstract

Kinesin-1 is a dimeric motor that transports cargo along microtubules, taking 8.2-nm steps in a hand-over-hand fashion. The ATP hydrolysis cycles of its two heads are maintained out of phase by a series of gating mechanisms, which lead to processive runs averaging ~1 μm. A key structural element for inter-head coordination is the neck linker (NL), which connects the heads to the stalk. To examine the role of the NL in regulating stepping, we investigated NL mutants of various lengths using single-molecule optical trapping and bulk fluorescence approaches in the context of a general framework for gating. Our results show that, although inter-head tension enhances motor velocity, it is crucial neither for inter-head coordination nor for rapid rear-head release. Furthermore, cysteine-light mutants do not produce wild-type motility under load. We conclude that kinesin-1 is primarily front-head gated, and that NL length is tuned to enhance unidirectional processivity and velocity.

Original languageEnglish (US)
Article numbere07403
Pages (from-to)1-44
Number of pages44
JournaleLife
Volume2015
Issue number4
DOIs
StatePublished - Apr 22 2015

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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    Andreasson, J. O. L., Milic, B., Chen, G. Y., Guydosh, N. R., Hancock, W. O., & Block, S. M. (2015). Examining kinesin processivity within a general gating framework. eLife, 2015(4), 1-44. [e07403]. https://doi.org/10.7554/eLife.07403