title: Nucleotide-dependent behavior of single molecules of cytoplasmic dynein on microtubules in vitro.
abstract: We visualized the nucleotide-dependent behavior of single molecules of mammalian native cytoplasmic dynein using fragments of dynactin p150 with or without its N-terminal microtubule binding domain. The results indicate that the binding affinity of dynein for microtubules is high in AMP-PNP, middle in ADP or no nucleotide, and low in ADP.Pi conditions. It is also demonstrated that the microtubule binding domain of dynactin p150 maintains the association of dynein with microtubules without altering the motile property of dynein in the weak binding state. In addition, we observed bidirectional movement of dynein in the presence of ATP as well as in ADP/Vi condition, suggesting that the bidirectional movement is driven by diffusion rather than active transport.
authors.affiliation: Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Tokyo, Japan.
authors.forename: Michi
authors.initials: M
authors.lastname: Miura
authors.forename: Aiko
authors.initials: A
authors.lastname: Matsubara
authors.forename: Takuya
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Masaki
authors.initials: M
authors.lastname: Edamatsu
authors.forename: Yoko Y
authors.initials: YY
authors.lastname: Toyoshima
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism; D056445:Cytoplasmic Dyneins / Q000378:metabolism*; D004058:Diffusion; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D009711:Nucleotides; C035135:periodate-oxidized ADP; D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D056445:Cytoplasmic Dyneins; D004398:Dyneins