Oscillatory movement of a dynein-microtubule complex crosslinked with DNA origami
Shimaa A Abdellatef,
Hisashi Tadakuma,
Kangmin Yan,
Takashi Fujiwara,
Kodai Fukumoto,
Yuichi Kondo,
Hiroko Takazaki,
Rofia Boudria,
Takuo Yasunaga,
Hideo Higuchi,
Keiko Hirose
Affiliations
Shimaa A Abdellatef
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan; Research Center for Functional Materials, National Institute for Materials Science, Tsukuba, Japan
Hisashi Tadakuma
Institute for Protein Research, Osaka University, Osaka, Japan; SLST and Gene Editing Center, ShanghaiTech University, Shanghai, China
Kangmin Yan
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan
Takashi Fujiwara
Graduate School of Science, The University of Tokyo, Tokyo, Japan
Kodai Fukumoto
Institute for Protein Research, Osaka University, Osaka, Japan
Yuichi Kondo
Graduate School of Science, The University of Tokyo, Tokyo, Japan
Hiroko Takazaki
Institute for Protein Research, Osaka University, Osaka, Japan; Kyushu Institute of Technology, Fukuoka, Japan
Rofia Boudria
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan; Institut Pasteur, Paris, France
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.
