The Kinesin-5 Chemomechanical Cycle Is Dominated by a Two-heads-bound State

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2016 Jul 13

PMID 27402829

Citations 21

Authors

Geng-Yuan Chen

Keith J Mickolajczyk

William O Hancock

Affiliations

Soon will be listed here.

Abstract

Single-molecule microscopy and stopped-flow kinetics assays were carried out to understand the microtubule polymerase activity of kinesin-5 (Eg5). Four lines of evidence argue that the motor primarily resides in a two-heads-bound (2HB) state. First, upon microtubule binding, dimeric Eg5 releases both bound ADPs. Second, microtubule dissociation in saturating ADP is 20-fold slower for the dimer than for the monomer. Third, ATP-triggered mant-ADP release is 5-fold faster than the stepping rate. Fourth, ATP binding is relatively fast when the motor is locked in a 2HB state. Shortening the neck-linker does not facilitate rear-head detachment, suggesting a minimal role for rear-head-gating. This 2HB state may enable Eg5 to stabilize incoming tubulin at the growing microtubule plus-end. The finding that slowly hydrolyzable ATP analogs trigger slower nucleotide release than ATP suggests that ATP hydrolysis in the bound head precedes stepping by the tethered head, leading to a mechanochemical cycle in which processivity is determined by the race between unbinding of the bound head and attachment of the tethered head.

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References

Jiang W, Stock M, Li X, Hackney D . Influence of the kinesin neck domain on dimerization and ATPase kinetics. J Biol Chem. 1997; 272(12):7626-32. DOI: 10.1074/jbc.272.12.7626. View

Shimamoto Y, Forth S, Kapoor T . Measuring Pushing and Braking Forces Generated by Ensembles of Kinesin-5 Crosslinking Two Microtubules. Dev Cell. 2015; 34(6):669-81. PMC: 4604754. DOI: 10.1016/j.devcel.2015.08.017. View

Scholey J, Nithianantham S, Scholey J, Al-Bassam J . Structural basis for the assembly of the mitotic motor Kinesin-5 into bipolar tetramers. Elife. 2014; 3:e02217. PMC: 3978770. DOI: 10.7554/eLife.02217. View

Chen Y, Hancock W . Kinesin-5 is a microtubule polymerase. Nat Commun. 2015; 6:8160. PMC: 4600729. DOI: 10.1038/ncomms9160. View

Myers K, Baas P . Kinesin-5 regulates the growth of the axon by acting as a brake on its microtubule array. J Cell Biol. 2007; 178(6):1081-91. PMC: 2064629. DOI: 10.1083/jcb.200702074. View

Behnke-Parks W, Vendome J, Honig B, Maliga Z, Moores C, Rosenfeld S . Loop L5 acts as a conformational latch in the mitotic kinesin Eg5. J Biol Chem. 2010; 286(7):5242-53. PMC: 3037637. DOI: 10.1074/jbc.M110.192930. View

Cochran J, Sontag C, Maliga Z, Kapoor T, Correia J, Gilbert S . Mechanistic analysis of the mitotic kinesin Eg5. J Biol Chem. 2004; 279(37):38861-70. PMC: 1356567. DOI: 10.1074/jbc.M404203200. View

Ruhnow F, Zwicker D, Diez S . Tracking single particles and elongated filaments with nanometer precision. Biophys J. 2011; 100(11):2820-8. PMC: 3117161. DOI: 10.1016/j.bpj.2011.04.023. View

Valentine M, Block S . Force and premature binding of ADP can regulate the processivity of individual Eg5 dimers. Biophys J. 2009; 97(6):1671-7. PMC: 2749793. DOI: 10.1016/j.bpj.2009.07.013. View

10.

Lebel P, Basu A, Oberstrass F, Tretter E, Bryant Z . Gold rotor bead tracking for high-speed measurements of DNA twist, torque and extension. Nat Methods. 2014; 11(4):456-62. PMC: 4211898. DOI: 10.1038/nmeth.2854. View

11.

Hariharan V, Hancock W . Insights into the Mechanical Properties of the Kinesin Neck Linker Domain from Sequence Analysis and Molecular Dynamics Simulations. Cell Mol Bioeng. 2011; 2(2):177-189. PMC: 3085455. DOI: 10.1007/s12195-009-0059-5. View

12.

Muretta J, Jun Y, Gross S, Major J, Thomas D, Rosenfeld S . The structural kinetics of switch-1 and the neck linker explain the functions of kinesin-1 and Eg5. Proc Natl Acad Sci U S A. 2015; 112(48):E6606-13. PMC: 4672802. DOI: 10.1073/pnas.1512305112. View

13.

Cochran J, Gilbert S . ATPase mechanism of Eg5 in the absence of microtubules: insight into microtubule activation and allosteric inhibition by monastrol. Biochemistry. 2005; 44(50):16633-48. PMC: 2270472. DOI: 10.1021/bi051724w. View

14.

Rosenfeld S, Xing J, Jefferson G, King P . Docking and rolling, a model of how the mitotic motor Eg5 works. J Biol Chem. 2005; 280(42):35684-95. DOI: 10.1074/jbc.M506561200. View

15.

Shastry S, Hancock W . Interhead tension determines processivity across diverse N-terminal kinesins. Proc Natl Acad Sci U S A. 2011; 108(39):16253-8. PMC: 3182723. DOI: 10.1073/pnas.1102628108. View

16.

Roostalu J, Hentrich C, Bieling P, Telley I, Schiebel E, Surrey T . Directional switching of the kinesin Cin8 through motor coupling. Science. 2011; 332(6025):94-9. DOI: 10.1126/science.1199945. View

17.

Hackney D . The tethered motor domain of a kinesin-microtubule complex catalyzes reversible synthesis of bound ATP. Proc Natl Acad Sci U S A. 2005; 102(51):18338-43. PMC: 1317912. DOI: 10.1073/pnas.0505288102. View

18.

Skoufias D, DeBonis S, Saoudi Y, Lebeau L, Crevel I, Cross R . S-trityl-L-cysteine is a reversible, tight binding inhibitor of the human kinesin Eg5 that specifically blocks mitotic progression. J Biol Chem. 2006; 281(26):17559-69. DOI: 10.1074/jbc.M511735200. View

19.

Kapitein L, Peterman E, Kwok B, Kim J, Kapoor T, Schmidt C . The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks. Nature. 2005; 435(7038):114-8. DOI: 10.1038/nature03503. View

20.

Kaan H, Hackney D, Kozielski F . The structure of the kinesin-1 motor-tail complex reveals the mechanism of autoinhibition. Science. 2011; 333(6044):883-5. PMC: 3339660. DOI: 10.1126/science.1204824. View