title: Lis1 restricts the conformational changes in cytoplasmic dynein on microtubules.
abstract: Cytoplasmic dynein is a microtubule-based motor protein that transports intracellular cargo and performs various functions during cell division. We previously reported that Lis1 suppressed dynein motility on microtubules in an idling state. Recently, a model showed that Lis1 prevents the ATPase domain of dynein from transmitting a detachment signal to its microtubule-binding domain. However, conformational information on dynein is limited. We used electron microscopy to investigate the conformation of dynein and nucleotide-induced conformational changes on microtubules. The conformation of dynein differed depending on the presence or absence of a nucleotide. In the presence of the nucleotide ADP-vanadate, dynein displayed an extended form on microtubules (extended form), whereas in the absence of a nucleotide, dynein lay along microtubules (compact form). This conformational change reflects chemomechanical coupling in dynein walking on microtubules. We also found that Lis1 fixed the conformation of dynein in the compact form regardless of the nucleotide condition. Removal of the Lis1 dimerization motif abolished Lis1-dependent fixation of dynein in the compact form. This suggests that the idling state of dynein on microtubules induced by Lis1 occurs through the Lis1-dependent arrest of dynein chemomechanical coupling.
authors.affiliation: Department of Genetic Disease Research, Osaka City University Graduate School of Medicine, Asahi-machi 1-4-3 Abeno, Osaka 545-8585, Japan.
authors.forename: Shiori
authors.initials: S
authors.lastname: Toba
authors.affiliation: Department of Bioscience and Bioinformatics, Faculty of Computer Science and Systems Engineering, Kyushu Institute of Technology, Kawazu 680-4, Iizuka, Fukuoka 820-8502, Japan JST-SENTAN, 4-1-8, Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Koyasako
authors.affiliation: Department of Bioscience and Bioinformatics, Faculty of Computer Science and Systems Engineering, Kyushu Institute of Technology, Kawazu 680-4, Iizuka, Fukuoka 820-8502, Japan JST-SENTAN, 4-1-8, Honcho, Kawaguchi, Saitama 332-0012, Japan JST-CREST, 4-1-8, Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Takuo
authors.initials: T
authors.lastname: Yasunaga
authors.affiliation: Department of Genetic Disease Research, Osaka City University Graduate School of Medicine, Asahi-machi 1-4-3 Abeno, Osaka 545-8585, Japan shinjih@med.osaka-cu.ac.jp.
authors.forename: Shinji
authors.initials: S
authors.lastname: Hirotsune
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D020285:Cryoelectron Microscopy / Q000379:methods*; D056445:Cytoplasmic Dyneins / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013552:Swine; D014638:Vanadates / Q000378:metabolism
chemical_list: D008869:Microtubule-Associated Proteins; D009711:Nucleotides; D014638:Vanadates; D056445:Cytoplasmic Dyneins
keywords: cryo-electron microscopy; electron microscopy; lissencephaly; microtubule; motor protein; single particle analysis