TY - JOUR
T1 - Single-Molecule Analysis of Dynein Processivity and Stepping Behavior
AU - Reck-Peterson, Samara L.
AU - Yildiz, Ahmet
AU - Carter, Andrew P.
AU - Gennerich, Arne
AU - Zhang, Nan
AU - Vale, Ronald D.
N1 - Funding Information:
The authors wish to thank Nicole Mahoney for early contributions, Nico Stuurman and Adam Douglass for advice on microscopy and image analysis, members of the Vale lab and Andres Leschziner for stimulating discussions, and Roberto Albanese for media preparation. We thank Jacob Kerssemakers and Marileen Dogterom for generously providing their step-finding program ahead of publication. This work has been supported by the National Institutes of Health (P01-AR42895 [R.D.V.] and F32-GM67403-02 [S.R.-P.]), the Jane Coffin Childs Foundation (A.G. and A.P.C.), the German Research Foundation (GE 1609/1 [A.G.]), the Agouron Institute (A.P.C.), and the Howard Hughes Medical Institute.
PY - 2006/7/28
Y1 - 2006/7/28
N2 - Cytoplasmic dynein, the 1.2 MDa motor driving minus-end-directed motility, has been reported to move processively along microtubules, but its mechanism of motility remains poorly understood. Here, using S. cerevisiae to produce recombinant dynein with a chemically controlled dimerization switch, we show by structural and single-molecule analysis that processivity requires two dynein motor domains but not dynein's tail domain or any associated subunits. Dynein advances most frequently in 8 nm steps, although longer as well as side and backward steps are observed. Individual motor domains show a different stepping pattern, which is best explained by the two motor domains shuffling in an alternating manner between rear and forward positions. Our results suggest that cytoplasmic dynein moves processively through the coordination of its two motor domains, but its variable step size and direction suggest a considerable diffusional component to its step, which differs from Kinesin-1 and is more akin to myosin VI.
AB - Cytoplasmic dynein, the 1.2 MDa motor driving minus-end-directed motility, has been reported to move processively along microtubules, but its mechanism of motility remains poorly understood. Here, using S. cerevisiae to produce recombinant dynein with a chemically controlled dimerization switch, we show by structural and single-molecule analysis that processivity requires two dynein motor domains but not dynein's tail domain or any associated subunits. Dynein advances most frequently in 8 nm steps, although longer as well as side and backward steps are observed. Individual motor domains show a different stepping pattern, which is best explained by the two motor domains shuffling in an alternating manner between rear and forward positions. Our results suggest that cytoplasmic dynein moves processively through the coordination of its two motor domains, but its variable step size and direction suggest a considerable diffusional component to its step, which differs from Kinesin-1 and is more akin to myosin VI.
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U2 - 10.1016/j.cell.2006.05.046
DO - 10.1016/j.cell.2006.05.046
M3 - Article
C2 - 16873064
AN - SCOPUS:33746253688
SN - 0092-8674
VL - 126
SP - 335
EP - 348
JO - Cell
JF - Cell
IS - 2
ER -