Cytoplasmic dynein is not a conventional processive motor.
Cytoplasmic dynein is a microtubule-based molecular motor with a multitude of functions from cell division to organelle transport. Cargo transport is often achieved as a co-complex with dynactin and it is believed that this co-complex enhances the processive translocation of cargo along the microtubule tracks (King and Schroer, 2000; Culver-Hanlon et al., 2006). Single molecule studies have revealed that dynein on its own can also act as a processive motor (Reck-Peterson et al., 2006; Toba et al., 2006). However, these studies did not allow the detection of a non-processive motor function. Previous studies based on the transport of vesicles or liposomes indicated that processive transport could only be achieved by an ensemble of motor molecules (Schroer & Sheetz, 1991; Wang and Sheetz, 2000; Muresan et al., 2001). Here we use the three bead dumbbell assay to show for the first time, that cytoplasmic dynein is a non-processive motor at low ATP concentrations. Processivity can be restored even in the absence of dynactin by increasing the ATP concentration to 100muM. We propose that an altered occupancy of the different ATP binding sites (AAA1-4) acts as a modulator between processive and non-processive stepping.
Walter
Wilhelm J
WJ
Medical School Hannover, Inst. Molec. & Cell Physiol., Carl-Neuberg Str. 1, 30625 Hannover, Germany.
Brenner
Bernhard
B
Steffen
Walter
W
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001692:Biological Transport / Q000502:physiology; D056445:Cytoplasmic Dyneins / Q000737:chemistry / Q000378:metabolism*; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D020409:Molecular Motor Proteins / Q000737:chemistry / Q000378:metabolism*; D052898:Optical Tweezers; D011485:Protein Binding; D013552:Swine; D014404:Tubulin / Q000378:metabolism
D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D020409:Molecular Motor Proteins; D014404:Tubulin; D000255:Adenosine Triphosphate; D056445:Cytoplasmic Dyneins