Motor domain phosphorylation modulates kinesin-1 transport

Hannah A. DeBerg, Benjamin H. Blehm, Janet Sheung, Andrew R. Thompson, Carol S. Bookwalter, Seyed F. Torabi, Trina A. Schroer, Christopher L. Berger, Yi Lu, Kathleen M. Trybus, Paul R. Selvin

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23 Scopus citations

Abstract

Disruptions in microtubule motor transport are associated with a variety of neurodegenerative diseases. Post-translational modification of the cargo-binding domain of the light and heavy chains of kinesin has been shown to regulate transport, but less is known about how modifications of the motor domain affect transport. Here we report on the effects of phosphorylation of a mammalian kinesin motor domain by the kinase JNK3 at a conserved serine residue (Ser-175 in the B isoform and Ser-176 in the A and C isoforms). Phosphorylation of this residue has been implicated in Huntington disease, but the mechanism by which Ser-175 phosphorylation affects transport is unclear. The ATPase, microtubule-binding affinity, and processivity are unchanged between a phosphomimetic S175D and a nonphosphorylatable S175A construct. However, we find that application of force differentiates between the two. Placement of negative charge at Ser-175, through phosphorylation or mutation, leads to a lower stall force and decreased velocity under a load of 1 piconewton or greater. Sedimentation velocity experiments also show that addition of a negative charge at Ser-175 favors the autoinhibited conformation of kinesin. These observations imply that when cargo is transported by both dynein and phosphorylated kinesin, a common occurrence in the cell, there may be a bias that favors motion toward the minus-end of microtubules. Such bias could be used to tune transport in healthy cells when properly regulated but contribute to a disease state when misregulated.

Original languageEnglish (US)
Pages (from-to)32612-32621
Number of pages10
JournalJournal of Biological Chemistry
Volume288
Issue number45
DOIs
StatePublished - Nov 8 2013

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    DeBerg, H. A., Blehm, B. H., Sheung, J., Thompson, A. R., Bookwalter, C. S., Torabi, S. F., Schroer, T. A., Berger, C. L., Lu, Y., Trybus, K. M., & Selvin, P. R. (2013). Motor domain phosphorylation modulates kinesin-1 transport. Journal of Biological Chemistry, 288(45), 32612-32621. https://doi.org/10.1074/jbc.M113.515510