|Entry||Database: PDB / ID: 5hnw|
|Title||Structural basis of backwards motion in kinesin-14: minus-end directed nKn664 in the AMPPNP state|
|Keywords||TRANSPORT PROTEIN / kinesin / kinesin-14 / microtubule / ATPase|
|Function / homology||Tubulin / Alpha tubulin / Kinesin motor domain / Tubulin subunits alpha, beta, and gamma signature. / Tubulin C-terminal domain / Kinesin motor domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ, C-terminal domain superfamily / Kinesin motor domain superfamily / Tubulin/FtsZ, GTPase domain superfamily ...Tubulin / Alpha tubulin / Kinesin motor domain / Tubulin subunits alpha, beta, and gamma signature. / Tubulin C-terminal domain / Kinesin motor domain / Tubulin/FtsZ family, GTPase domain / Tubulin/FtsZ, C-terminal domain superfamily / Kinesin motor domain superfamily / Tubulin/FtsZ, GTPase domain superfamily / Kinesin motor domain signature. / Beta tubulin / Tubulin/FtsZ, GTPase domain / Kinesin-like protein / P-loop containing nucleoside triphosphate hydrolase / Tubulin, C-terminal / Kinesin motor domain, conserved site / Tubulin/FtsZ, 2-layer sandwich domain / Tubulin, conserved site / Tubulin-beta mRNA autoregulation signal. / Kinesin motor domain profile. / Tubulin/FtsZ, C-terminal / RHO GTPases activate IQGAPs / Kinesins / Carboxyterminal post-translational modifications of tubulin / The role of GTSE1 in G2/M progression after G2 checkpoint / Mitotic Prometaphase / COPI-independent Golgi-to-ER retrograde traffic / COPI-dependent Golgi-to-ER retrograde traffic / COPI-mediated anterograde transport / RHO GTPases Activate Formins / Intraflagellar transport / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Hedgehog 'off' state / Recycling pathway of L1 / Recruitment of NuMA to mitotic centrosomes / HSP90 chaperone cycle for steroid hormone receptors (SHR) / Resolution of Sister Chromatid Cohesion / Separation of Sister Chromatids / MHC class II antigen presentation / Beta tubulin, autoregulation binding site / spindle assembly involved in female meiosis / minus-end directed microtubule sliding / spindle assembly involved in meiosis / regulation of mitotic spindle elongation / distributive segregation / mitotic spindle microtubule / mitotic spindle elongation / meiotic spindle organization / ATP-dependent microtubule motor activity, minus-end-directed / microtubule bundle formation / regulation of mitotic spindle assembly / positive regulation of axon guidance / meiotic spindle / mitotic centrosome separation / kinesin complex / microtubule motor activity / mRNA transport / spindle organization / mitotic spindle assembly / cytoplasmic microtubule / cellular response to interleukin-4 / microtubule-based movement / microtubule-based process / mitotic spindle organization / chromosome segregation / mitotic spindle / structural constituent of cytoskeleton / microtubule cytoskeleton organization / neuron migration / microtubule cytoskeleton / spindle / double-stranded RNA binding / mitotic cell cycle / microtubule / microtubule binding / ATPase activity / GTPase activity / centrosome / cell division / GTP binding / ubiquitin protein ligase binding / protein heterodimerization activity / protein homodimerization activity / ATP binding / nucleus / cytosol / cytoplasm / Protein claret segregational / Tubulin alpha-1B chain / Tubulin beta-2B chain|
Function and homology information
|Specimen source||Drosophila melanogaster (fruit fly)|
Rattus norvegicus (Norway rat)
Bos taurus (cattle)
|Method||ELECTRON MICROSCOPY / helical reconstruction / cryo EM / 6.6 Å resolution|
|Authors||Shigematsu, H. / Yokoyama, T. / Kikkawa, M. / Shirouzu, M. / Nitta, R.|
|Citation||Journal: Structure / Year: 2016|
Title: Structural Basis of Backwards Motion in Kinesin-1-Kinesin-14 Chimera: Implication for Kinesin-14 Motility.
Authors: Masahiko Yamagishi / Hideki Shigematsu / Takeshi Yokoyama / Masahide Kikkawa / Mitsuhiro Sugawa / Mari Aoki / Mikako Shirouzu / Junichiro Yajima / Ryo Nitta
Abstract: Kinesin-14 is a unique minus-end-directed microtubule-based motor. A swinging motion of a class-specific N-terminal neck helix has been proposed to produce minus-end directionality. However, it is ...Kinesin-14 is a unique minus-end-directed microtubule-based motor. A swinging motion of a class-specific N-terminal neck helix has been proposed to produce minus-end directionality. However, it is unclear how swinging of the neck helix is driven by ATP hydrolysis utilizing the highly conserved catalytic core among all kinesins. Here, using a motility assay, we show that in addition to the neck helix, the conserved five residues at the C-terminal region in kinesin-14, namely the neck mimic, are necessary to give kinesin-1 an ability to reverse its directionality toward the minus end of microtubules. Our structural analyses further demonstrate that the C-terminal neck mimic, in cooperation with conformational changes in the catalytic core during ATP binding, forms a kinesin-14 bundle with the N-terminal neck helix to swing toward the minus end of microtubules. Thus, the neck mimic plays a crucial role in coupling the chemical ATPase reaction with the mechanical cycle to produce the minus-end-directed motility of kinesin-14.
SummaryFull reportAbout validation report
|Date||Deposition: Jan 19, 2016 / Release: Aug 10, 2016|
|Structure viewer||Molecule: |
Downloads & links
A: Tubulin alpha-1B chain
B: Tubulin beta-2B chain
K: Protein claret segregational,KINESIN HEAVY CHAIN ISOFORM 5C
-Protein/peptide , 3 types, 3 molecules A
|#1: Protein/peptide|| |
Mass: 49976.043 Da / Num. of mol.: 1 / Source: (natural) Bos taurus (cattle) / References: UniProt: P81947
|#2: Protein/peptide|| |
Mass: 49776.570 Da / Num. of mol.: 1 / Source: (natural) Bos taurus (cattle) / References: UniProt: Q6B856
|#3: Protein/peptide|| |
Mass: 41620.250 Da / Num. of mol.: 1 / Fragment: UNP RESIDUES 325-348, 664-700
Source: (gene. exp.) Drosophila melanogaster (fruit fly), (gene. exp.) Rattus norvegicus (Norway rat)
Gene: ncd, CA(ND), CG7831 / Production host: Escherichia coli (E. coli) / References: UniProt: P20480
-Non-polymers , 5 types, 6 molecules
|#4: Chemical||#5: Chemical|| ChemComp-GTP / ||#6: Chemical|| ChemComp-GDP / ||#7: Chemical|| ChemComp-TA1 / ||#8: Chemical|| ChemComp-ANP / |
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: FILAMENT / Reconstruction method: helical reconstruction|
|Buffer solution||pH: 6.8|
|Specimen||Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Talos Arctica / Image courtesy: FEI Company
|Microscopy||Microscope model: FEI TECNAI ARCTICA|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy|
|Image recording||Electron dose: 30 e/Å2 / Film or detector model: FEI FALCON II (4k x 4k)|
|CTF correction||Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|Helical symmerty||Angular rotation/subunit: -25.719666 deg. / Axial rise/subunit: 8.648646 Å / Axial symmetry: C1|
|3D reconstruction||Resolution: 6.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 229516 / Details: High-resolution noise substitution was performed / Symmetry type: HELICAL|
|Atomic model building||Ref protocol: FLEXIBLE FIT|
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