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Force Generation of KIF1C Is Impaired by Pathogenic Mutations

25 Pages Posted: 4 Apr 2022 Publication Status: Published

See all articles by Nida Siddiqui

Nida Siddiqui

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Daniel Roth

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Algirdas Toleikis

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Alexander J. Zwetsloot

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Robert A. Cross

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Anne Straube

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

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Abstract

Intracellular transport is essential for neuronal function and survival. The most effective plus end-directed neuronal transporter is the kinesin-3 KIF1C, which transports large secretory vesicles and endosomes. Mutations in KIF1C cause hereditary spastic paraplegia and cerebellar dysfunction in human patients. In contrast to other kinesin-3s, KIF1C is a stable dimer and a highly processive motor in its native state. Here we establish a baseline for the single molecule mechanics of Kif1C. We show that full length KIF1C molecules can processively step against the load of an optical trap and reach average stall forces of 3.7 pN. Compared to kinesin-1, KIF1C has a higher propensity to slip backwards under load, which results in a lower maximal single molecule force. However, KIF1C remains attached to the microtubule while slipping backwards and re-engages quickly consistent with its super-processivity. Two pathogenic mutations P176L and R169W that cause hereditary spastic paraplegia in humans maintain fast, processive single molecule motility in vitro, but with decreased run length and slightly increased unloaded velocity compared to the wildtype motor. Under load in an optical trap, force generation by these mutants is severely reduced. In cells, the same mutants are impaired in producing sufficient force to efficiently relocate organelles. Our results show how its mechanics supports KIF1C’s role as an intracellular transporter and explain how pathogenic mutations at the microtubule-binding interface of KIF1C impair the cellular function of these long-distance transporters and result in neuronal disease.

Keywords: molecular motor, kinesin, KIF1C, optical trap, hereditary spastic paraplegia, intracellular transport, single molecule force, microtubule binding, kinesin-3

Suggested Citation

Siddiqui, Nida and Roth, Daniel and Toleikis, Algirdas and Zwetsloot, Alexander J. and Cross, Robert A. and Straube, Anne, Force Generation of KIF1C Is Impaired by Pathogenic Mutations. Available at SSRN: https://ssrn.com/abstract=4075230 or http://dx.doi.org/10.2139/ssrn.4075230
This version of the paper has not been formally peer reviewed.

Nida Siddiqui

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Coventry
United Kingdom

Daniel Roth

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Coventry
United Kingdom

Algirdas Toleikis

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Coventry
United Kingdom

Alexander J. Zwetsloot

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Coventry
United Kingdom

Robert A. Cross

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences

Coventry
United Kingdom

Anne Straube (Contact Author)

University of Warwick - Centre for Mechanochemical Cell Biology & Division of Biomedical Sciences ( email )

Coventry
United Kingdom

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