title: Kinesin-2 KIF3AC and KIF3AB Can Drive Long-Range Transport along Microtubules.
abstract: Mammalian KIF3AC is classified as a heterotrimeric kinesin-2 that is best known for organelle transport in neurons, yet in vitro studies to characterize its single molecule behavior are lacking. The results presented show that a KIF3AC motor that includes the native helix α7 sequence for coiled-coil formation is highly processive with run lengths of ∼1.23 μm and matching those exhibited by conventional kinesin-1. This result was unexpected because KIF3AC exhibits the canonical kinesin-2 neck-linker sequence that has been reported to be responsible for shorter run lengths observed for another heterotrimeric kinesin-2, KIF3AB. However, KIF3AB with its native neck linker and helix α7 is also highly processive with run lengths of ∼1.62 μm and exceeding those of KIF3AC and kinesin-1. Loop L11, a component of the microtubule-motor interface and implicated in activating ADP release upon microtubule collision, is significantly extended in KIF3C as compared with other kinesins. A KIF3AC encoding a truncation in KIF3C loop L11 (KIF3ACΔL11) exhibited longer run lengths at ∼1.55 μm than wild-type KIF3AC and were more similar to KIF3AB run lengths, suggesting that L11 also contributes to tuning motor processivity. The steady-state ATPase results show that shortening L11 does not alter kcat, consistent with the observation that single molecule velocities are not affected by this truncation. However, shortening loop L11 of KIF3C significantly increases the microtubule affinity of KIF3ACΔL11, revealing another structural and mechanistic property that can modulate processivity. The results presented provide new, to our knowledge, insights to understand structure-function relationships governing processivity and a better understanding of the potential of KIF3AC for long-distance transport in neurons.
authors.affiliation: Department of Biological Sciences and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Guzik-Lendrum
authors.affiliation: Department of Biochemistry, University of Wisconsin, Madison, Wisconsin.
authors.forename: Katherine C
authors.initials: KC
authors.lastname: Rank
authors.affiliation: Department of Biological Sciences and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York.
authors.forename: Brandon M
authors.initials: BM
authors.lastname: Bensel
authors.affiliation: Department of Biochemistry, University of Wisconsin, Madison, Wisconsin.
authors.forename: Keenan C
authors.initials: KC
authors.lastname: Taylor
authors.affiliation: Department of Biochemistry, University of Wisconsin, Madison, Wisconsin. Electronic address: ivan_rayment@biochem.wisc.edu.
authors.forename: Ivan
authors.initials: I
authors.lastname: Rayment
authors.affiliation: Department of Biological Sciences and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York. Electronic address: sgilbert@rpi.edu.
authors.forename: Susan P
authors.initials: SP
authors.lastname: Gilbert
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D019281:Dimerization; D004926:Escherichia coli; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D045663:Quantum Dots; D017385:Sequence Homology; D014741:Video Recording
chemical_list: C490202:Kif3a protein, mouse; C490672:Kif3b protein, mouse; C484676:Kif3c protein, mouse; D000251:Adenosine Triphosphatases; D016547:Kinesins