Oscillatory movement of a dynein-microtubule complex crosslinked with DNA origami

eLife

Hideo Higuchi¹  Hisashi Tadakuma²  Takashi Fujiwara³  Kodai Fukumoto⁴  Shimaa A Abdellatef⁴  Keiko Hirose⁴  Rofia Boudria⁵  Yuichi Kondo⁵  Takuo Yasunaga⁶  Hiroko Takazaki⁶  Kangmin Yan⁶ 

[1] Kyushu Institute of Technology, Fukuoka, Japan;Research Center for Functional Materials, National Institute for Materials Science, Tsukuba, Japan;SLST and Gene Editing Center, ShanghaiTech University, Shanghai, China;Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan;Graduate School of Science, The University of Tokyo, Tokyo, Japan;Institute for Protein Research, Osaka University, Osaka, Japan;

关键词: motor proteins;    dynein;    flagella;    DNA origami;    molecular biophysics;   

DOI  :  10.7554/eLife.76357

来源: DOAJ

【 摘 要 】

Bending of cilia and flagella occurs when axonemal dynein molecules on one side of the axoneme produce force and move toward the microtubule (MT) minus end. These dyneins are then pulled back when the axoneme bends in the other direction, meaning oscillatory back and forth movement of dynein during repetitive bending of cilia/flagella. There are various factors that may regulate the dynein activity, e.g. the nexin-dynein regulatory complex, radial spokes, and central apparatus. In order to understand the basic mechanism of dynein’s oscillatory movement, we constructed a simple model system composed of MTs, outer-arm dyneins, and crosslinks between the MTs made of DNA origami. Electron microscopy (EM) showed pairs of parallel MTs crossbridged by patches of regularly arranged dynein molecules bound in two different orientations, depending on which of the MTs their tails bind to. The oppositely oriented dyneins are expected to produce opposing forces when the pair of MTs have the same polarity. Optical trapping experiments showed that the dynein-MT-DNA-origami complex actually oscillates back and forth after photolysis of caged ATP. Intriguingly, the complex, when held at one end, showed repetitive bending motions. The results show that a simple system composed of ensembles of oppositely oriented dyneins, MTs, and inter-MT crosslinkers, without any additional regulatory structures, has an intrinsic ability to cause oscillation and repetitive bending motions.

【 授权许可】

Unknown