KIF3A Accelerates KIF3C within the Kinesin-2 Heterodimer to Generate Symmetrical Phosphate Release Rates for each Processive Step

Heterodimeric KIF3AC is a mammalian kinesin-2 that is highly expressed in the central nervous system and is associated with vesicles in neurons. KIF3AC is an intriguing member of the kinesin-2 family because the intrinsic kinetics of KIF3A and KIF3C when expressed as homodimers and analyzed in vitro are distinctively different from each other. For example, the single-molecule velocities of the engineered homodimers KIF3AA and KIF3CC are 293 nm/s and 7.5 nm/s, respectively, whereas KIF3AC has a velocity of 186 nm/s. These results led us to hypothesize that heterodimerization alters the intrinsic catalytic properties of the two heads, and an earlier computational analysis predicted that processive steps would alternate between a fast step for KIF3A followed by a slow step for KIF3C resulting in asymmetric stepping during a processive run. To test this hypothesis directly, we measured the presteady-state kinetics of phosphate release for KIF3AC, KIF3AA, and KIF3CC followed by computational modeling of the KIF3AC phosphate release transients. The results reveal that KIF3A and KIF3C retain their intrinsic ATP binding and hydrolysis kinetics. Yet within KIF3AC, KIF3A activates the rate of phosphate release for KIF3C such that the coupled steps of phosphate release and dissociation from the microtubule become more similar for KIF3A and KIF3C. These coupled steps are the rate-limiting transition for the ATPase cycle suggesting that within KIF3AC, the stepping kinetics are similar for each head during the processive run. The model is also consistent with either KIF3A or KIF3C ending the processive run.

Reference

S.M. Quinn, T. Vargason, N. Pokhrel, E. Antony, J. Hahn, and S.P. Gilbert. "KIF3A Accelerates KIF3C within the Kinesin-2 Heterodimer to Generate Symmetrical Phosphate Release Rates for each Processive Step"

Journal of Biological Chemistry, In Press (2020)