Slow axonemal dynein e facilitates the motility of faster dynein c.
We highly purified the Chlamydomonas inner-arm dyneins e and c, considered to be single-headed subspecies. These two dyneins reside side-by-side along the peripheral doublet microtubules of the flagellum. Electron microscopic observations and single particle analysis showed that the head domains of these two dyneins were similar, whereas the tail domain of dynein e was short and bent in contrast to the straight tail of dynein c. The ATPase activities, both basal and microtubule-stimulated, of dynein e (kcat = 0.27 s(-1) and kcat,MT = 1.09 s(-1), respectively) were lower than those of dynein c (kcat = 1.75 s(-1) and kcat,MT = 2.03 s(-1), respectively). From in vitro motility assays, the apparent velocity of microtubule translocation by dynein e was found to be slow (Vap = 1.2 ± 0.1 μm/s) and appeared independent of the surface density of the motors, whereas dynein c was very fast (Vmax = 15.8 ± 1.5 μm/s) and highly sensitive to decreases in the surface density (Vmin = 2.2 ± 0.7 μm/s). Dynein e was expected to be a processive motor, since the relationship between the microtubule landing rate and the surface density of dynein e fitted well with first-power dependence. To obtain insight into the in vivo roles of dynein e, we measured the sliding velocity of microtubules driven by a mixture of dynein e and c at various ratios. The microtubule translocation by the fast dynein c became even faster in the presence of the slow dynein e, which could be explained by assuming that dynein e does not retard motility of faster dyneins. In flagella, dynein e likely acts as a facilitator by holding adjacent microtubules to aid dynein c's power stroke.
National Institute of Information and Communications Technology (NICT), Advanced ICT Research Institute, Hyogo, Japan.
Youské
Y
Shimizu
National Institute of Information and Communications Technology (NICT), Advanced ICT Research Institute, Hyogo, Japan.
Hitoshi
H
Sakakibara
National Institute of Information and Communications Technology (NICT), Advanced ICT Research Institute, Hyogo, Japan.
Hiroaki
H
Kojima
National Institute of Information and Communications Technology (NICT), Advanced ICT Research Institute, Hyogo, Japan; Japan Science and Technology Agency (JST), Core Research for Evolutionary Science and Technology (CREST), Tokyo, Japan. Electronic address: oiwa@nict.go.jp.
Kazuhiro
K
Oiwa
D000251:Adenosine Triphosphatases / Q000378:metabolism; D056444:Axonemal Dyneins / Q000235:genetics / Q000378:metabolism*; D002696:Chlamydomonas / Q000166:cytology / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism; D007700:Kinetics; D008870:Microtubules / Q000378:metabolism; D009068:Movement; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport
D010940:Plant Proteins; D000251:Adenosine Triphosphatases; D056444:Axonemal Dyneins