title: A flipped ion pair at the dynein-microtubule interface is critical for dynein motility and ATPase activation.
abstract: Dynein is a motor protein that moves on microtubules (MTs) using the energy of adenosine triphosphate (ATP) hydrolysis. To understand its motility mechanism, it is crucial to know how the signal of MT binding is transmitted to the ATPase domain to enhance ATP hydrolysis. However, the molecular basis of signal transmission at the dynein-MT interface remains unclear. Scanning mutagenesis of tubulin identified two residues in α-tubulin, R403 and E416, that are critical for ATPase activation and directional movement of dynein. Electron cryomicroscopy and biochemical analyses revealed that these residues form salt bridges with the residues in the dynein MT-binding domain (MTBD) that work in concert to induce registry change in the stalk coiled coil and activate the ATPase. The R403-E3390 salt bridge functions as a switch for this mechanism because of its reversed charge relative to other residues at the interface. This study unveils the structural basis for coupling between MT binding and ATPase activation and implicates the MTBD in the control of directional movement.
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Uchimura
authors.affiliation: Graduate School of Frontier Biosciences and Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Kawaguchi, Saitama 332-0012, Japan Quantitative Biology Center, Institute of Physical and Chemical Research, Suita, Osaka 565-0871, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Fujii
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Takazaki
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Ayukawa
authors.affiliation: Graduate School of Frontier Biosciences and Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Nishikawa
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Itsushi
authors.initials: I
authors.lastname: Minoura
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: You
authors.initials: Y
authors.lastname: Hachikubo
authors.affiliation: Graduate School of Frontier Biosciences and Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan Department of Macromolecular Science, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
authors.forename: Genji
authors.initials: G
authors.lastname: Kurisu
authors.affiliation: Research Institute for Science and Engineering, Waseda University, Toshima-ku, Tokyo 171-0033, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sutoh
authors.affiliation: Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Kawaguchi, Saitama 332-0012, Japan Department of Macromolecular Science, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan Department of Frontier Bioscience, Faculty of Bioscience and Applied Chemistry, Hosei University, Koganei, Tokyo 184-8584, Japan.
authors.forename: Takahide
authors.initials: T
authors.lastname: Kon
authors.affiliation: Graduate School of Frontier Biosciences and Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan Quantitative Biology Center, Institute of Physical and Chemical Research, Suita, Osaka 565-0871, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Namba
authors.affiliation: Laboratory for Molecular Biophysics, RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan emuto2@brain.riken.jp.
authors.forename: Etsuko
authors.initials: E
authors.lastname: Muto
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D020285:Cryoelectron Microscopy; D004023:Dictyostelium; D004398:Dyneins / Q000737:chemistry* / Q000648:ultrastructure; D004789:Enzyme Activation; D008870:Microtubules / Q000737:chemistry* / Q000648:ultrastructure; D008958:Models, Molecular; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D020836:Protein Structure, Quaternary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000648:ultrastructure; D034421:Sus scrofa
chemical_list: D015800:Protozoan Proteins; D004398:Dyneins