The Structure of the Kinesin-1 Motor-Tail Complex Reveals the Mechanism of Autoinhibition

A tail domain autoinhibits a dimeric kinesin by preventing relative movement of the two motor domains. When not transporting cargo, kinesin-1 is autoinhibited by binding of a tail region to the motor domains, but the mechanism of inhibition is unclear. We report the crystal structure of a motor domain dimer in complex with its tail domain at 2.2 angstroms and compare it with a structure of the motor domain alone at 2.7 angstroms. These structures indicate that neither an induced conformational change nor steric blocking is the cause of inhibition. Instead, the tail cross-links the motor domains at a second position, in addition to the coiled coil. This “double lockdown,” by cross-linking at two positions, prevents the movement of the motor domains that is needed to undock the neck linker and release adenosine diphosphate. This autoinhibition mechanism could extend to some other kinesins.

[1]  Michael P. Sheetz,et al.  Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility , 1985, Cell.

[2]  D. Hackney,et al.  Half-site inhibition of dimeric kinesin head domains by monomeric tail domains. , 2009, Biochemistry.

[3]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[4]  K. Henrick,et al.  Inference of macromolecular assemblies from crystalline state. , 2007, Journal of molecular biology.

[5]  Martin Karplus,et al.  Force generation in kinesin hinges on cover-neck bundle formation. , 2008, Structure.

[6]  An allosteric transition trapped in an intermediate state of a new kinesin-inhibitor complex. , 2009, The Biochemical journal.

[7]  N. Hirokawa,et al.  Kinesin and dynein superfamily proteins and the mechanism of organelle transport. , 1998, Science.

[8]  Y. Wong,et al.  Kinesin’s light chains inhibit the head- and microtubule-binding activity of its tail , 2010, Proceedings of the National Academy of Sciences.

[9]  F. Kull,et al.  Kinesin: switch I & II and the motor mechanism. , 2002, Journal of cell science.

[10]  K. Verhey,et al.  Traffic control: regulation of kinesin motors , 2009, Nature Reviews Molecular Cell Biology.

[11]  E. Mandelkow,et al.  The Crystal Structure of Dimeric Kinesin and Implications for Microtubule-Dependent Motility , 1997, Cell.

[12]  Hee-Won Park,et al.  Rotation of the stalk/neck and one head in a new crystal structure of the kinesin motor protein, Ncd , 2003, The EMBO journal.

[13]  Albert Sickmann,et al.  Feedback of the Kinesin-1 Neck-linker Position on the Catalytic Site* , 2006, Journal of Biological Chemistry.

[14]  S. Seiler,et al.  Cargo binding and regulatory sites in the tail of fungal conventional kinesin , 2000, Nature Cell Biology.

[15]  F. Kull,et al.  Kinesins at a glance , 2010, Journal of Cell Science.

[16]  Vincent B. Chen,et al.  Correspondence e-mail: , 2000 .

[17]  Roger Cooke,et al.  A structural change in the kinesin motor protein that drives motility , 1999, Nature.

[18]  Kenneth H Downing,et al.  The kinesin-1 motor protein is regulated by a direct interaction of its head and tail , 2008, Proceedings of the National Academy of Sciences.

[19]  Kenneth H Downing,et al.  An atomic-level mechanism for activation of the kinesin molecular motors , 2010, Proceedings of the National Academy of Sciences.

[20]  F. Kozielski,et al.  The crystal structure of the minus-end-directed microtubule motor protein ncd reveals variable dimer conformations. , 1999, Structure.

[21]  Ronald D. Vale,et al.  Crystal structure of the kinesin motor domain reveals a structural similarity to myosin , 1996, Nature.

[22]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[23]  Ting Huang,et al.  Formation of the Compact Confomer of Kinesin Requires a COOH-terminal Heavy Chain Domain and Inhibits Microtubule-stimulated ATPase Activity* , 1999, The Journal of Biological Chemistry.

[24]  Randy J. Read,et al.  Phaser crystallographic software , 2007, Journal of applied crystallography.

[25]  P. Evans,et al.  Scaling and assessment of data quality. , 2006, Acta crystallographica. Section D, Biological crystallography.

[26]  K. Kobashi,et al.  Catalytic oxidation of sulfhydryl groups by o-phenanthroline copper complex. , 1968, Biochimica et biophysica acta.

[27]  D. Hackney,et al.  Interaction of mant-adenosine nucleotides and magnesium with kinesin. , 1998, Biochemistry.

[28]  Ahmad S. Khalil,et al.  Kinesin's cover-neck bundle folds forward to generate force , 2008, Proceedings of the National Academy of Sciences.

[29]  M. F. Stock,et al.  Kinesin’s IAK tail domain inhibits initial microtubule-stimulated ADP release , 2000, Nature Cell Biology.

[30]  Noboru Yumoto,et al.  Mechanism of tail-mediated inhibition of kinesin activities studied using synthetic peptides. , 2006, Biochemical and biophysical research communications.

[31]  Wei Jiang,et al.  Influence of the Kinesin Neck Domain on Dimerization and ATPase Kinetics* , 1997, The Journal of Biological Chemistry.

[32]  M. F. Stock,et al.  Kinesin tail domains and Mg2+ directly inhibit release of ADP from head domains in the absence of microtubules. , 2008, Biochemistry.

[33]  M. Seeger,et al.  Microtubule-associated Protein-like Binding of the Kinesin-1 Tail to Microtubules* , 2010, The Journal of Biological Chemistry.

[34]  Ronald D. Vale,et al.  Direction determination in the minus-end-directed kinesin motor ncd , 1998, Nature.