A Tubulin Binding Switch Underlies Kip3/Kinesin-8 Depolymerase Activity

• Published in: Developmental cell Vol. 42; no. 1; pp. 37 - 51.e8
• Main Authors: Arellano-Santoyo, Hugo, Geyer, Elisabeth A., Stokasimov, Ema, Chen, Geng-Yuan, Su, Xiaolei, Hancock, William, Rice, Luke M., Pellman, David
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 10.07.2017
• Subjects: Adenosine Triphosphate - metabolism; Animals; Biocatalysis; depolymerization; Hydrolysis; Kinesin - chemistry; Kinesin - metabolism; kinesins; microtubule associated proteins; microtubule dynamics; Microtubules - metabolism; Models, Biological; Models, Molecular; Mutant Proteins - metabolism; Polymerization; Protein Binding; Protein Structure, Secondary; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - metabolism; spindle scaling; Sus scrofa; Tubulin - metabolism; microtubule associated proteins; spindle scaling; depolymerization; kinesins; microtubule dynamics
• DOI: 10.1016/j.devcel.2017.06.011
• ISSN: 1534-5807

Kinesin-8 motors regulate the size of microtubule structures, using length-dependent accumulation at the plus end to preferentially disassemble long microtubules. Despite extensive study, the kinesin-8 depolymerase mechanism remains under debate. Here, we provide evidence for an alternative, tubulin curvature-sensing model of microtubule depolymerization by the budding yeast kinesin-8, Kip3. Kinesin-8/Kip3 uses ATP hydrolysis, like other kinesins, for stepping on the microtubule lattice, but at the plus end Kip3 undergoes a switch: its ATPase activity is suppressed when it binds tightly to the curved conformation of tubulin. This prolongs plus-end binding, stabilizes protofilament curvature, and ultimately promotes microtubule disassembly. The tubulin curvature-sensing model is supported by our identification of Kip3 structural elements necessary and sufficient for plus-end binding and depolymerase activity, as well as by the identification of an α-tubulin residue specifically required for the Kip3-curved tubulin interaction. Together, these findings elucidate a major regulatory mechanism controlling the size of cellular microtubule structures. •A tubulin binding switch enables length-dependent microtubule disassembly by Kip3•Tubulin binding suppresses Kip3 ATPase activity, promoting depolymerase activity•Loop11 sequence in Kip3 is necessary and sufficient for curved tubulin binding•Kip3 disassembly-resistant microtubules support the two-state binding switch model Kinesin-8s promote length-dependent microtubule disassembly. Arellano and colleagues define a mechanism for Kip3/kinesin-8 depolymerization that involves a tubulin curvature-sensing binding switch. They propose that high affinity curved tubulin binding, which is accompanied by suppressed ATPse activity, enables its depolymerase activity. This Kip3 binding switch is thus central to Kip3’s ability to selectively trim long microtubules, narrowing the length distribution of microtubules in cells.