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- EMDB-2395: MuB is an AAA+ ATPase that forms helical filaments to control tar... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2395 | |||||||||
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Title | MuB is an AAA+ ATPase that forms helical filaments to control target selection for DNA transposition | |||||||||
![]() | Reconstruction of MuB filament without DNA | |||||||||
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![]() | AAA+ ATPase / DNA transposition / Mu phage / nucleoprotein filament / symmetry mismatch | |||||||||
Function / homology | ![]() phosphorelay signal transduction system / sequence-specific DNA binding / regulation of DNA-templated transcription / ATP hydrolysis activity / ATP binding / identical protein binding / metal ion binding Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | helical reconstruction / cryo EM / Resolution: 18.0 Å | |||||||||
![]() | Mizuno N / Dramicanin M / Mizuuchi M / Adam J / Wang Y / Han YW / Yang W / Steven AC / Mizuuchi K / Ramon-Maiques S | |||||||||
![]() | ![]() Title: MuB is an AAA+ ATPase that forms helical filaments to control target selection for DNA transposition. Authors: Naoko Mizuno / Marija Dramićanin / Michiyo Mizuuchi / Julia Adam / Yi Wang / Yong-Woon Han / Wei Yang / Alasdair C Steven / Kiyoshi Mizuuchi / Santiago Ramón-Maiques / ![]() Abstract: MuB is an ATP-dependent nonspecific DNA-binding protein that regulates the activity of the MuA transposase and captures target DNA for transposition. Mechanistic understanding of MuB function has ...MuB is an ATP-dependent nonspecific DNA-binding protein that regulates the activity of the MuA transposase and captures target DNA for transposition. Mechanistic understanding of MuB function has previously been hindered by MuB's poor solubility. Here we combine bioinformatic, mutagenic, biochemical, and electron microscopic analyses to unmask the structure and function of MuB. We demonstrate that MuB is an ATPase associated with diverse cellular activities (AAA+ ATPase) and forms ATP-dependent filaments with or without DNA. We also identify critical residues for MuB's ATPase, DNA binding, protein polymerization, and MuA interaction activities. Using single-particle electron microscopy, we show that MuB assembles into a helical filament, which binds the DNA in the axial channel. The helical parameters of the MuB filament do not match those of the coated DNA. Despite this protein-DNA symmetry mismatch, MuB does not deform the DNA duplex. These findings, together with the influence of MuB filament size on strand-transfer efficiency, lead to a model in which MuB-imposed symmetry transiently deforms the DNA at the boundary of the MuB filament and results in a bent DNA favored by MuA for transposition. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 1.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 11 KB 11 KB | Display Display | ![]() |
Images | ![]() | 146.9 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 4bs1MC ![]() 2398C ![]() 2400C ![]() 4bt0C ![]() 4bt1C M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Reconstruction of MuB filament without DNA | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : MuB filament without DNA
Entire | Name: MuB filament without DNA |
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Components |
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-Supramolecule #1000: MuB filament without DNA
Supramolecule | Name: MuB filament without DNA / type: sample / ID: 1000 / Oligomeric state: helical filament / Number unique components: 1 |
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Molecular weight | Experimental: 35 KDa / Theoretical: 35 KDa |
-Macromolecule #1: MuB AAA+ ATPase
Macromolecule | Name: MuB AAA+ ATPase / type: protein_or_peptide / ID: 1 / Oligomeric state: helical assembly / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() |
Molecular weight | Experimental: 35 KDa / Theoretical: 35 KDa |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | helical reconstruction |
Aggregation state | filament |
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Sample preparation
Concentration | 0.07 mg/mL |
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Buffer | pH: 8 Details: 30 mM TrisHCl pH 8.0, 0.15 M KCl, 5mM MgCl2, 1mM DTT, 1 mM ATP or ATP-gamma-S |
Grid | Details: 300 mesh quantifoil R2/2 |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 90 K / Instrument: FEI VITROBOT MARK II / Method: Blot for 5 seconds before plunging |
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Electron microscopy
Microscope | FEI/PHILIPS CM200FEG |
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Temperature | Min: 80 K / Max: 85 K / Average: 82 K |
Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 100,000 times magnification |
Date | Apr 21, 2008 |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 1000 (2k x 2k) / Number real images: 129 / Average electron dose: 15 e/Å2 |
Tilt angle min | 0 |
Tilt angle max | 0 |
Electron beam | Acceleration voltage: 120 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 38000 |
Sample stage | Specimen holder: side entry / Specimen holder model: GATAN LIQUID NITROGEN |
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Image processing
Details | IHRSR |
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Final reconstruction | Applied symmetry - Helical parameters - Δz: 9.01 Å Applied symmetry - Helical parameters - Δ&Phi: 66.2 ° Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 18.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: spider, bsoft, eman |
CTF correction | Details: phase-flipping |