Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Published Online December 18, 2003
Science DOI: 10.1126/science.1093753

Reports

Submitted on November 17, 2003
Accepted on December 10, 2003

Kinesin Walks Hand-Over-Hand

Ahmet Yildiz 1, Michio Tomishige 2, Ronald D. Vale 3, Paul R. Selvin 4*

1 Center for Biophysics and Computational Biology, University of Illinois, Urbana-Champaign, IL 61801, USA.
2 The Howard Hughes Medical Institute and the Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94107, USA; Current address: Department of Applied Physics, The University of Tokyo, Tokyo 113-8656, Japan.
3 The Howard Hughes Medical Institute and the Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94107, USA.
4 Center for Biophysics and Computational Biology and Physics Department, University of Illinois, Urbana-Champaign, IL 61801, USA.

* To whom correspondence should be addressed. E-mail: selvin{at}uiuc.edu.

Kinesin is a processive motor that takes 8.3 nm center-of-mass steps along microtubules for each ATP hydrolyzed. Whether kinesin moves by a "hand-over-hand" or an "inchworm" model has been controversial. We have labeled a single head of the kinesin dimer with a Cy3 fluorophore and localized the position of the dye within 2 nm before and after a step. We observe that single kinesin heads take 17.3 ± 3.3 nm steps. A kinetic analysis of the dwell times between steps shows that the 17 nm steps alternate with 0 nm steps. These results strongly support a hand-over-hand mechanism, and not an inchworm mechanism. In addition, our results suggest that kinesin is bound via both heads to the microtubule while waiting for ATP in between steps.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Interactions between Subunits in Heterodimeric Ncd Molecules.
E. Kocik, K. J. Skowronek, and A. A. Kasprzak (2009)
J. Biol. Chem. 284, 35735-35745
   Abstract »    Full Text »    PDF »
Kinesin's step dissected with single-motor FRET.
S. Verbrugge, Z. Lansky, and E. J. G. Peterman (2009)
PNAS 106, 17741-17746
   Abstract »    Full Text »    PDF »
Direct measurements of kinesin torsional properties reveal flexible domains and occasional stalk reversals during stepping.
B. Gutierrez-Medina, A. N. Fehr, and S. M. Block (2009)
PNAS 106, 17007-17012
   Abstract »    Full Text »    PDF »
Why kinesin is so processive.
E. Toprak, A. Yildiz, M. T. Hoffman, S. S. Rosenfeld, and P. R. Selvin (2009)
PNAS 106, 12717-12722
   Abstract »    Full Text »    PDF »
A mobile kinesin-head intermediate during the ATP-waiting state.
A. B. Asenjo and H. Sosa (2009)
PNAS 106, 5657-5662
   Abstract »    Full Text »    PDF »
Dynamics of the bacterial flagellar motor with multiple stators.
G. Meacci and Y. Tu (2009)
PNAS 106, 3746-3751
   Abstract »    Full Text »    PDF »
Regulatory ATPase Sites of Cytoplasmic Dynein Affect Processivity and Force Generation.
C. Cho, S. L. Reck-Peterson, and R. D. Vale (2008)
J. Biol. Chem. 283, 25839-25845
   Abstract »    Full Text »    PDF »
The role of microtubule movement in bidirectional organelle transport.
I. M. Kulic, A. E. X. Brown, H. Kim, C. Kural, B. Blehm, P. R. Selvin, P. C. Nelson, and V. I. Gelfand (2008)
PNAS 105, 10011-10016
   Abstract »    Full Text »    PDF »
Intracellular Transport and Kinesin Superfamily Proteins, KIFs: Structure, Function, and Dynamics.
N. Hirokawa and Y. Noda (2008)
Physiol Rev 88, 1089-1118
   Abstract »    Full Text »    PDF »
The mitotic kinesin CENP-E is a processive transport motor.
H. Yardimci, M. van Duffelen, Y. Mao, S. S. Rosenfeld, and P. R. Selvin (2008)
PNAS 105, 6016-6021
   Abstract »    Full Text »    PDF »
Single-molecule biophysics: at the interface of biology, physics and chemistry.
A. A Deniz, S. Mukhopadhyay, and E. A Lemke (2008)
J R Soc Interface 5, 15-45
   Abstract »    Full Text »    PDF »
Synergic mechanism and fabrication target for bipedal nanomotors.
Z. Wang (2007)
PNAS 104, 17921-17926
   Abstract »    Full Text »    PDF »
Mechanical control of the directional stepping dynamics of the kinesin motor.
C. Hyeon and J. N. Onuchic (2007)
PNAS 104, 17382-17387
   Abstract »    Full Text »    PDF »
Not So Lame After All: Kinesin Still Walks with a Hobbled Head.
N. R. Guydosh and S. M. Block (2007)
J. Gen. Physiol. 130, 441-444
   Full Text »    PDF »
Myosin V stepping mechanism.
G. Cappello, P. Pierobon, C. Symonds, L. Busoni, J. Christof, M. Gebhardt, M. Rief, and J. Prost (2007)
PNAS 104, 15328-15333
   Abstract »    Full Text »    PDF »
From the Cover: Detection of fractional steps in cargo movement by the collective operation of kinesin-1 motors.
C. Leduc, F. Ruhnow, J. Howard, and S. Diez (2007)
PNAS 104, 10847-10852
   Abstract »    Full Text »    PDF »
Myosin V Walks by Lever Action and Brownian Motion.
K. Shiroguchi and K. Kinosita Jr. (2007)
Science 316, 1208-1212
   Abstract »    Full Text »    PDF »
An ATP Gate Controls Tubulin Binding by the Tethered Head of Kinesin-1.
M. C. Alonso, D. R. Drummond, S. Kain, J. Hoeng, L. Amos, and R. A. Cross (2007)
Science 316, 120-123
   Abstract »    Full Text »    PDF »
Tracking melanosomes inside a cell to study molecular motors and their interaction.
C. Kural, A. S. Serpinskaya, Y.-H. Chou, R. D. Goldman, V. I. Gelfand, and P. R. Selvin (2007)
PNAS 104, 5378-5382
   Abstract »    Full Text »    PDF »
Extending the absorbing boundary method to fit dwell-time distributions of molecular motors with complex kinetic pathways.
J.-C. Liao, J. A. Spudich, D. Parker, and S. L. Delp (2007)
PNAS 104, 3171-3176
   Abstract »    Full Text »    PDF »
Use of plasmon coupling to reveal the dynamics of DNA bending and cleavage by single EcoRV restriction enzymes.
B. M. Reinhard, S. Sheikholeslami, A. Mastroianni, A. P. Alivisatos, and J. Liphardt (2007)
PNAS 104, 2667-2672
   Abstract »    Full Text »    PDF »
Sustained Activity of Calcium Release-activated Calcium Channels Requires Translocation of Mitochondria to the Plasma Membrane.
A. Quintana, E. C. Schwarz, C. Schwindling, P. Lipp, L. Kaestner, and M. Hoth (2006)
J. Biol. Chem. 281, 40302-40309
   Abstract »    Full Text »    PDF »
Feedback of the Kinesin-1 Neck-linker Position on the Catalytic Site.
K. Hahlen, B. Ebbing, J. Reinders, J. Mergler, A. Sickmann, and G. Woehlke (2006)
J. Biol. Chem. 281, 18868-18877
   Abstract »    Full Text »    PDF »
Backsteps induced by nucleotide analogs suggest the front head of kinesin is gated by strain.
N. R. Guydosh and S. M. Block (2006)
PNAS 103, 8054-8059
   Abstract »    Full Text »    PDF »
On the hand-over-hand mechanism of kinesin.
Q. Shao and Y. Q. Gao (2006)
PNAS 103, 8072-8077
   Abstract »    Full Text »    PDF »
Defocused orientation and position imaging (DOPI) of myosin V.
E. Toprak, J. Enderlein, S. Syed, S. A. McKinney, R. G. Petschek, T. Ha, Y. E. Goldman, and P. R. Selvin (2006)
PNAS 103, 6495-6499
   Abstract »    Full Text »    PDF »
Overlapping hand-over-hand mechanism of single molecular motility of cytoplasmic dynein.
S. Toba, T. M. Watanabe, L. Yamaguchi-Okimoto, Y. Y. Toyoshima, and H. Higuchi (2006)
PNAS 103, 5741-5745
   Abstract »    Full Text »    PDF »
Kinesin crouches to sprint but resists pushing.
M. E. Fisher and Y. C. Kim (2005)
PNAS 102, 16209-16214
   Abstract »    Full Text »    PDF »
Alternating Site ATPase Pathway of Rat Conventional Kinesin.
S. D. Auerbach and K. A. Johnson (2005)
J. Biol. Chem. 280, 37048-37060
   Abstract »    Full Text »    PDF »
From The Cover: Measurement of the contributions of 1D and 3D pathways to the translocation of a protein along DNA.
D. M. Gowers, G. G. Wilson, and S. E. Halford (2005)
PNAS 102, 15883-15888
   Abstract »    Full Text »    PDF »
Microoxen: Microorganisms to move microscale loads.
D. B. Weibel, P. Garstecki, D. Ryan, W. R. DiLuzio, M. Mayer, J. E. Seto, and G. M. Whitesides (2005)
PNAS 102, 11963-11967
   Abstract »    Full Text »    PDF »
Monte Carlo modeling of single-molecule cytoplasmic dynein.
M. P. Singh, R. Mallik, S. P. Gross, and C. C. Yu (2005)
PNAS 102, 12059-12064
   Abstract »    Full Text »    PDF »
Kinesin and Dynein Move a Peroxisome in Vivo: A Tug-of-War or Coordinated Movement?.
C. Kural, H. Kim, S. Syed, G. Goshima, V. I. Gelfand, and P. R. Selvin (2005)
Science 308, 1469-1472
   Abstract »    Full Text »    PDF »
Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time.
L. S. Churchman, Z. Okten, R. S. Rock, J. F. Dawson, and J. A. Spudich (2005)
PNAS 102, 1419-1423
   Abstract »    Full Text »    PDF »
From the Cover: Cooperative extraction of membrane nanotubes by molecular motors.
C. Leduc, O. Campas, K. B. Zeldovich, A. Roux, P. Jolimaitre, L. Bourel-Bonnet, B. Goud, J.-F. Joanny, P. Bassereau, and J. Prost (2004)
PNAS 101, 17096-17101
   Abstract »    Full Text »    PDF »
Mechanistic Analysis of the Mitotic Kinesin Eg5.
J. C. Cochran, C. A. Sontag, Z. Maliga, T. M. Kapoor, J. J. Correia, and S. P. Gilbert (2004)
J. Biol. Chem. 279, 38861-38870
   Abstract »    Full Text »    PDF »
Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave.
A. Sarkar, R. B. Robertson, and J. M. Fernandez (2004)
PNAS 101, 12882-12886
   Abstract »    Full Text »    PDF »
Nanometer-localized multiple single-molecule fluorescence microscopy.
X. Qu, D. Wu, L. Mets, and N. F. Scherer (2004)
PNAS 101, 11298-11303
   Abstract »    Full Text »    PDF »
Expression of several cytoskeletal proteins in ovine cerebral arteries: developmental and functional considerations.
Y. Zhao, H. Xiao, W. Long, W. J. Pearce, and L. D. Longo (2004)
J. Physiol. 558, 623-632
   Abstract »    Full Text »    PDF »
How do site-specific DNA-binding proteins find their targets?.
S. E. Halford and J. F. Marko (2004)
Nucleic Acids Res. 32, 3040-3052
   Abstract »    Full Text »    PDF »
Single-molecule high-resolution imaging with photobleaching.
M. P. Gordon, T. Ha, and P. R. Selvin (2004)
PNAS 101, 6462-6465
   Abstract »    Full Text »    PDF »
Coordination between Motor Domains in Processive Kinesins.
E. P. Sablin and R. J. Fletterick (2004)
J. Biol. Chem. 279, 15707-15710
   Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)