Monte Carlo modeling of single-molecule cytoplasmic dynein.

Molecular motors are responsible for active transport and organization in the cell, underlying an enormous number of crucial biological processes. Dynein is more complicated in its structure and function than other motors. Recent experiments have found that, unlike other motors, dynein can take different size steps along microtubules depending on load and ATP concentration. We use Monte Carlo simulations to model the molecular motor function of cytoplasmic dynein at the single-molecule level. The theory relates dynein's enzymatic properties to its mechanical force production. Our simulations reproduce the main features of recent single-molecule experiments that found a discrete distribution of dynein step sizes, depending on load and ATP concentration. The model reproduces the large steps found experimentally under high ATP and no load by assuming that the ATP binding affinities at the secondary sites decrease as the number of ATP bound to these sites increases. Additionally, to capture the essential features of the step-size distribution at very low ATP concentration and no load, the ATP hydrolysis of the primary site must be dramatically reduced when none of the secondary sites have ATP bound to them. We make testable predictions that should guide future experiments related to dynein function.

[1]  R. Vale,et al.  Comparison of the motile and enzymatic properties of two microtubule minus-end-directed motors, ncd and cytoplasmic dynein. , 1995, Biochemistry.

[2]  T. Elston,et al.  A model for the oscillatory motion of single dynein molecules. , 2005, Journal of theoretical biology.

[3]  M L Yarmush,et al.  Dissociation kinetics of antigen-antibody interactions: studies on a panel of anti-albumin monoclonal antibodies. , 1989, Molecular immunology.

[4]  Mark J. Schnitzer,et al.  Kinesin hydrolyses one ATP per 8-nm step , 1997, Nature.

[5]  Y. Toyoshima,et al.  A Single-headed Recombinant Fragment of Dictyostelium Cytoplasmic Dynein Can Drive the Robust Sliding of Microtubules* , 2004, Journal of Biological Chemistry.

[6]  C. Bustamante,et al.  The mechanochemistry of molecular motors. , 2000, Biophysical journal.

[7]  A. Mehta,et al.  Myosin-V stepping kinetics: a molecular model for processivity. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Ronald D. Vale,et al.  Aaa Proteins , 2000, The Journal of cell biology.

[9]  Steven M. Block,et al.  Force and velocity measured for single kinesin molecules , 1994, Cell.

[10]  A. Silvanovich,et al.  The third P-loop domain in cytoplasmic dynein heavy chain is essential for dynein motor function and ATP-sensitive microtubule binding. , 2003, Molecular biology of the cell.

[11]  R. Vale,et al.  Kinesin Walks Hand-Over-Hand , 2004, Science.

[12]  Mark J. Schnitzer,et al.  Single kinesin molecules studied with a molecular force clamp , 1999, Nature.

[13]  Samara L. Reck-Peterson,et al.  Molecular dissection of the roles of nucleotide binding and hydrolysis in dynein's AAA domains in Saccharomyces cerevisiae. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[14]  G. Oster,et al.  Reverse engineering a protein: the mechanochemistry of ATP synthase. , 2000, Biochimica et biophysica acta.

[15]  J. Rothman,et al.  N-ethylmaleimide-sensitive fusion protein: a trimeric ATPase whose hydrolysis of ATP is required for membrane fusion , 1994, The Journal of cell biology.

[16]  A B Kolomeisky,et al.  The force exerted by a molecular motor. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  A. Kolomeisky,et al.  Simple mechanochemistry describes the dynamics of kinesin molecules , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  S. King,et al.  Design and regulation of the AAA+ microtubule motor dynein. , 2004, Journal of structural biology.

[19]  B. C. Carter,et al.  Cytoplasmic dynein functions as a gear in response to load , 2004, Nature.

[20]  S. Burgess,et al.  The structure of dynein-c by negative stain electron microscopy. , 2004, Journal of structural biology.

[21]  S. Leibler,et al.  Porters versus rowers: a unified stochastic model of motor proteins , 1993, The Journal of cell biology.

[22]  E. Holzbaur,et al.  ADP release is rate limiting in steady-state turnover by the dynein adenosinetriphosphatase. , 1989, Biochemistry.

[23]  S. Burgess,et al.  Dynein structure and power stroke , 2003, Nature.

[24]  Toshio Yanagida,et al.  Dynein arms are oscillating force generators , 1998, Nature.

[25]  J. Gelles,et al.  Coupling of kinesin steps to ATP hydrolysis , 1997, Nature.

[26]  Matthias Rief,et al.  Myosin-V is a processive actin-based motor , 1999, Nature.

[27]  M. Sheetz,et al.  Tracking kinesin-driven movements with nanometre-scale precision , 1988, Nature.

[28]  G. Mocz,et al.  Probing the nucleotide binding sites of axonemal dynein with the fluorescent nucleotide analogue 2'(3')-O-(-N-Methylanthraniloyl)-adenosine 5'-triphosphate. , 1998, Biochemistry.

[29]  D. Hackney,et al.  The kinetic cycles of myosin, kinesin, and dynein. , 1996, Annual review of physiology.

[30]  P. Selvin,et al.  Nanometer localization of single green fluorescent proteins: evidence that myosin V walks hand-over-hand via telemark configuration. , 2004, Biophysical journal.

[31]  Jimin Wang Nucleotide-dependent domain motions within rings of the RecA/AAA(+) superfamily. , 2004, Journal of structural biology.

[32]  Kazuo Sutoh,et al.  Distinct functions of nucleotide-binding/hydrolysis sites in the four AAA modules of cytoplasmic dynein. , 2004, Biochemistry.

[33]  K. Johnson The pathway of ATP hydrolysis by dynein. Kinetics of a presteady state phosphate burst. , 1983, The Journal of biological chemistry.

[34]  M. Welte,et al.  Bidirectional Transport along Microtubules , 2004, Current Biology.

[35]  T. Hays,et al.  Evidence for cooperative interactions between the two motor domains of cytoplasmic dynein , 1999, Current Biology.

[36]  G. Mocz,et al.  Model for the motor component of dynein heavy chain based on homology to the AAA family of oligomeric ATPases. , 2001, Structure.