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- EMDB-23345: Cryo-EM structure of Mycobacterium smegmatis Lhr helicase C-termi... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-23345 | |||||||||
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Title | Cryo-EM structure of Mycobacterium smegmatis Lhr helicase C-terminal domain | |||||||||
![]() | Cryo-EM structure of Mycobacterium smegmatis Lhr helicase C-terminal domain | |||||||||
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![]() | helicase / winged helix / HYDROLASE | |||||||||
Function / homology | ![]() hydrolase activity, acting on acid anhydrides, in phosphorus-containing anhydrides / nucleic acid binding / RNA helicase activity / RNA helicase / ATP binding Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
![]() | Wang J / Warren GM | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Oligomeric quaternary structure of Escherichia coli and Mycobacterium smegmatis Lhr helicases is nucleated by a novel C-terminal domain composed of five winged-helix modules. Authors: Garrett M Warren / Juncheng Wang / Dinshaw J Patel / Stewart Shuman / ![]() Abstract: Mycobacterium smegmatis Lhr (MsmLhr; 1507-aa) is the founder of a novel clade of bacterial helicases. MsmLhr consists of an N-terminal helicase domain (aa 1-856) with a distinctive tertiary structure ...Mycobacterium smegmatis Lhr (MsmLhr; 1507-aa) is the founder of a novel clade of bacterial helicases. MsmLhr consists of an N-terminal helicase domain (aa 1-856) with a distinctive tertiary structure (Lhr-Core) and a C-terminal domain (Lhr-CTD) of unknown structure. Here, we report that Escherichia coli Lhr (EcoLhr; 1538-aa) is an ATPase, translocase and ATP-dependent helicase. Like MsmLhr, EcoLhr translocates 3' to 5' on ssDNA and unwinds secondary structures en route, with RNA:DNA hybrid being preferred versus DNA:DNA duplex. The ATPase and translocase activities of EcoLhr inhere to its 877-aa Core domain. Full-length EcoLhr and MsmLhr have homo-oligomeric quaternary structures in solution, whereas their respective Core domains are monomers. The MsmLhr CTD per se is a homo-oligomer in solution. We employed cryo-EM to solve the structure of the CTD of full-length MsmLhr. The CTD protomer is composed of a series of five winged-helix (WH) modules and a β-barrel module. The CTD adopts a unique homo-tetrameric quaternary structure. A Lhr-CTD subdomain, comprising three tandem WH modules and the β-barrel, is structurally homologous to AlkZ, a bacterial DNA glycosylase that recognizes and excises inter-strand DNA crosslinks. This homology is noteworthy given that Lhr is induced in mycobacteria exposed to the inter-strand crosslinker mitomycin C. | |||||||||
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 | ![]() | 59.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 12 KB 12 KB | Display Display | ![]() |
Images | ![]() | 27.9 KB | ||
Filedesc metadata | ![]() | 6.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 568.9 KB | Display | ![]() |
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Full document | ![]() | 568.5 KB | Display | |
Data in XML | ![]() | 6 KB | Display | |
Data in CIF | ![]() | 6.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7lhlMC 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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Map
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Annotation | Cryo-EM structure of Mycobacterium smegmatis Lhr helicase C-terminal domain | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.064 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 : Tetramer structure of ATP-dependent DNA helicase Lhr C-terminal domain
Entire | Name: Tetramer structure of ATP-dependent DNA helicase Lhr C-terminal domain |
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Components |
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-Supramolecule #1: Tetramer structure of ATP-dependent DNA helicase Lhr C-terminal domain
Supramolecule | Name: Tetramer structure of ATP-dependent DNA helicase Lhr C-terminal domain type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 260 kDa/nm |
-Macromolecule #1: ATP-dependent DNA helicase
Macromolecule | Name: ATP-dependent DNA helicase / type: protein_or_peptide / ID: 1 / Number of copies: 4 / Enantiomer: LEVO / EC number: RNA helicase |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 162.009562 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: SMTTNGADPL GRFSALTREW FTTAFAAPTP AQADAWSAIS EGNNTLVIAP TGSGKTLAAF LWAIDRLADP AREPSQGTQV LYVSPLKAL AVDVERNLRT PLTGITRVAE RHGLPAPSIT VGVRSGDTPP NQRRAMIANP PDVLITTPES LFLMLTSAAR E TLTSVRTV ...String: SMTTNGADPL GRFSALTREW FTTAFAAPTP AQADAWSAIS EGNNTLVIAP TGSGKTLAAF LWAIDRLADP AREPSQGTQV LYVSPLKAL AVDVERNLRT PLTGITRVAE RHGLPAPSIT VGVRSGDTPP NQRRAMIANP PDVLITTPES LFLMLTSAAR E TLTSVRTV IVDEVHAVAA TKRGAHLALS LERLDQLLDT PAQRIGLSAT VRPPEEVARF LSGQAPTTIV CPPAAKTFDL SV QVPVPDM ANLDNNSIWP DVEERIVDLV EAHNSSIVFA NSRRLAERLT SRLNEIHAER SGIELPAGPN PEVGGGAPAH LMG SGQANG APPLLARAHH GSVSKEQRAQ VEDDLKSGRL RAVVATSSLE LGIDMGAVDL VIQVEAPPSV ASGLQRVGRA GHQV GEISQ GVLFPKHRTD LIGCAVTVQR MQTGDIETLR VPANPLDVLA QHTVAVAALE PVDADAWFDA VRRSAPFATL PRSAF EATL DLLSGKYPST EFAELRPRLV YDRDTGTLTA RPGAQRLAVT SGGAIPDRGM FTVYLASETE KPSRVGELDE EMVYES RPG DVISLGATSW RITEITHDRV LVIPAPGQPA RLPFWRGDSV GRPAELGAAV GAFTGELASL DRKAFDKRCQ KMGFAGY AT DNLHQLLREQ REATGVVPSD TTFVVERFRD ELGDWRVILH SPYGLRVHGP LALAVGRRLR ERYGIDEKPT ASDDGIIV R LPDSGDTPPG ADLFVFDADE IEPIVTAEVG GSALFASRFR ECAARALLLP RRHPGKRSPL WHQRQRAAQL LDIARKYPD FPIVLEAVRE CLQDVYDVPA LIELMHKIAQ RRLRIVEVET ATPSPFAASL LFGYVGAFMY EGDSPLAERR AAALALDTVL LSELLGRVE LRELLDPAVV ASTSAQLQHL TPERAARDAE GVADLLRLLG PLTEADIAQR CTADNIGAWL DGLHAAKRAL P VTYAGQTW WAAVEDIGLL RDGIGVPVPV GVPAAFTESA SDPLGDLIGR YARTRGPFTT EQTAARFGLG VRVASDVLSR MA VDGRLIR GEFAADLSGE QWCDAQVLKI LRRRSLAALR AQVEPVSTDA YARFLPSWQH VGSTNTTGVD GLATVIEQLA GVP IPASAV ESLVFPQRVR DYQPAMLDEL LASGEVMWSG AGQIGNGDGW VAFHLADTAP LTLTHGAEIE FTDTHRVILE TLGH GGAYF FRQLTDGTVE GTAGQELKQA LWELIWAGWV TGDTFAPVRA VLSGPRRSGA PAHRQRQRPP RLSRYSVAHA QTRGT DPTV SGRWSALPAA EPDSTVRAHF QAELLLGRHG VLTKGAVGAE GVPGGFATLY KVLSAFEDAG RCQRGYFVES LGGAQF AVA STVDRLRSYL DNVDPERPEY HAVVLAATDP ANPYGAALGW PTDSEAHRPG RKAGALVALV DGRLVWFLER GGRSLLS FG ADADAQRAAA GALTDLVSAG RIPSLLVERI NGVAVLDPDV DAERAVVQDA LLGAGLSRTP RGLRLR UniProtKB: ATP-dependent DNA helicase |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.3 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: UltrAuFoil / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 120 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 278 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 53.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 47262 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal magnification: 22500 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: OTHER |
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Final reconstruction | Number classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 101323 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD / Software - Name: RELION |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |