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- EMDB-15934: Cryo-EM structure of MLE in complex with ADP:AlF4 and SL7modUUC RNA -
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Open data
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Basic information
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Title | Cryo-EM structure of MLE in complex with ADP:AlF4 and SL7modUUC RNA | |||||||||
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![]() | RNA Helicase / Drosophila dosage compensation / RNA BINDING PROTEIN | |||||||||
Function / homology | ![]() RIP-mediated NFkB activation via ZBP1 / X chromosome located dosage compensation complex, transcription activating / dosage compensation complex assembly / 3'-5' DNA/RNA helicase activity / DEx/H-box helicases activate type I IFN and inflammatory cytokines production / male courtship behavior, veined wing generated song production / regulation of cytoplasmic translation / regulatory region RNA binding / PKR-mediated signaling / MSL complex ...RIP-mediated NFkB activation via ZBP1 / X chromosome located dosage compensation complex, transcription activating / dosage compensation complex assembly / 3'-5' DNA/RNA helicase activity / DEx/H-box helicases activate type I IFN and inflammatory cytokines production / male courtship behavior, veined wing generated song production / regulation of cytoplasmic translation / regulatory region RNA binding / PKR-mediated signaling / MSL complex / dosage compensation by hyperactivation of X chromosome / positive regulation of DNA topoisomerase (ATP-hydrolyzing) activity / sex-chromosome dosage compensation / 3'-5' RNA helicase activity / regulation of mRNA processing / axon extension / lncRNA binding / DNA duplex unwinding / 3'-5' DNA helicase activity / nuclear chromosome / positive regulation of heterochromatin formation / X chromosome / DNA helicase activity / helicase activity / determination of adult lifespan / double-stranded RNA binding / chromosome / double-stranded DNA binding / RNA helicase activity / RNA helicase / ribonucleoprotein complex / chromatin binding / chromatin / nucleolus / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / RNA binding / ATP binding / nucleus / cytosol Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.04 Å | |||||||||
![]() | Jagtap PKA / Hennig J | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis of RNA-induced autoregulation of the DExH-type RNA helicase maleless. Authors: Pravin Kumar Ankush Jagtap / Marisa Müller / Anna E Kiss / Andreas W Thomae / Karine Lapouge / Martin Beck / Peter B Becker / Janosch Hennig / ![]() Abstract: RNA unwinding by DExH-type helicases underlies most RNA metabolism and function. It remains unresolved if and how the basic unwinding reaction of helicases is regulated by auxiliary domains. We ...RNA unwinding by DExH-type helicases underlies most RNA metabolism and function. It remains unresolved if and how the basic unwinding reaction of helicases is regulated by auxiliary domains. We explored the interplay between the RecA and auxiliary domains of the RNA helicase maleless (MLE) from Drosophila using structural and functional studies. We discovered that MLE exists in a dsRNA-bound open conformation and that the auxiliary dsRBD2 domain aligns the substrate RNA with the accessible helicase tunnel. In an ATP-dependent manner, dsRBD2 associates with the helicase module, leading to tunnel closure around ssRNA. Furthermore, our structures provide a rationale for blunt-ended dsRNA unwinding and 3'-5' translocation by MLE. Structure-based MLE mutations confirm the functional relevance of our model for RNA unwinding. Our findings contribute to our understanding of the fundamental mechanics of auxiliary domains in DExH helicase MLE, which serves as a model for its human ortholog and potential therapeutic target, DHX9/RHA. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 60.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.3 KB 17.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 14.7 KB | Display | ![]() |
Images | ![]() | 69.9 KB | ||
Masks | ![]() | 125 MB | ![]() | |
Filedesc metadata | ![]() | 6.6 KB | ||
Others | ![]() ![]() | 116 MB 116 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8b9kMC ![]() 8b9gC ![]() 8b9iC ![]() 8b9jC ![]() 8b9lC ![]() 8pjbC ![]() 8pjjC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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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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Voxel size | X=Y=Z: 0.822 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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-Half map: #2
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Density Histograms |
-Half map: #1
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Density Histograms |
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Sample components
-Entire : MLE in complex with ADP:AlF4 and SL7modUUC RNA
Entire | Name: MLE in complex with ADP:AlF4 and SL7modUUC RNA |
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Components |
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-Supramolecule #1: MLE in complex with ADP:AlF4 and SL7modUUC RNA
Supramolecule | Name: MLE in complex with ADP:AlF4 and SL7modUUC RNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#2 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 149 KDa |
-Macromolecule #1: Dosage compensation regulator
Macromolecule | Name: Dosage compensation regulator / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA helicase |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 130.46507 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MDIKSFLYQF CAKSQIEPKF DIRQTGPKNR QRFLCEVRVE PNTYIGVGNS TNKKDAEKNA CRDFVNYLVR VGKLNTNDVP ADAGASGGG PRTGLEGAGM AGGSGQQKRV FDGQSGPQDL GEAYRPLNHD GGDGGNRYSV IDRIQEQRDM NEAEAFDVNA A IHGNWTIE ...String: MDIKSFLYQF CAKSQIEPKF DIRQTGPKNR QRFLCEVRVE PNTYIGVGNS TNKKDAEKNA CRDFVNYLVR VGKLNTNDVP ADAGASGGG PRTGLEGAGM AGGSGQQKRV FDGQSGPQDL GEAYRPLNHD GGDGGNRYSV IDRIQEQRDM NEAEAFDVNA A IHGNWTIE NAKERLNIYK QTNNIRDDYK YTPVGPEHAR SFLAELSIYV PALNRTVTAR ESGSNKKSAS KSCALSLVRQ LF HLNVIEP FSGTLKKKKD EQLKPYPVKL SPNLINKIDE VIKGLDLPVV NPRNIKIELD GPPIPLIVNL SRIDSSQQDG EKR QESSVI PWAPPQANWN TWHACNIDEG ELATTSIDDL SMDYERSLRD RRQNDNEYRQ FLEFREKLPI AAMRSEILTA INDN PVVII RGNTGCGKTT QIAQYILDDY ICSGQGGYAN IYVTQPRRIS AISVAERVAR ERCEQLGDTV GYSVRFESVF PRPYG AILF CTVGVLLRKL EAGLRGVSHI IVDEIHERDV NSDFLLVILR DMVDTYPDLH VILMSATIDT TKFSKYFGIC PVLEVP GRA FPVQQFFLED IIQMTDFVPS AESRRKRKEV EDEEQLLSED KDEAEINYNK VCEDKYSQKT RNAMAMLSES DVSFELL EA LLMHIKSKNI PGAILVFLPG WNLIFALMKF LQNTNIFGDT SQYQILPCHS QIPRDEQRKV FEPVPEGVTK IILSTNIA E TSITIDDIVF VIDICKARMK LFTSHNNLTS YATVWASKTN LEQRKGRAGR VRPGFCFTLC SRARFQALED NLTPEMFRT PLHEMALTIK LLRLGSIHHF LSKALEPPPV DAVIEAEVLL REMRCLDAND ELTPLGRLLA RLPIEPRLGK MMVLGAVFGC ADLMAIMAS YSSTFSEVFS LDIGQRRLAN HQKALSGTKC SDHVAMIVAS QMWRREKQRG EHMEARFCDW KGLQMSTMNV I WDAKQQLL DLLQQAGFPE ECMISHEVDE RIDGDDPVLD VSLALLCLGL YPNICVHKEK RKVLTTESKA ALLHKTSVNC SN LAVTFPY PFFVFGEKIR TRAVSCKQLS MVSPLQVILF GSRKIDLAAN NIVRVDNWLN FDIEPELAAK IGALKPALED LIT VACDNP SDILRLEEPY AQLVKVVKDL CVKSAGDFGL QRE UniProtKB: Dosage compensation regulator mle |
-Macromolecule #2: SL7modUUC
Macromolecule | Name: SL7modUUC / type: rna / ID: 2 / Number of copies: 1 |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 19.011131 KDa |
Sequence | String: GUGUAAAAUG UUGCUAGCAA AUAUAUAUGC UAGUAACGUU UUACGCCCUC UUUCUUUCUU |
-Macromolecule #3: ADENOSINE-5'-DIPHOSPHATE
Macromolecule | Name: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 3 / Number of copies: 1 / Formula: ADP |
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Molecular weight | Theoretical: 427.201 Da |
Chemical component information | ![]() ChemComp-ADP: |
-Macromolecule #4: TETRAFLUOROALUMINATE ION
Macromolecule | Name: TETRAFLUOROALUMINATE ION / type: ligand / ID: 4 / Number of copies: 1 / Formula: ALF |
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Molecular weight | Theoretical: 102.975 Da |
Chemical component information | ![]() ChemComp-ALF: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.612 mg/mL |
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Buffer | pH: 7.5 Details: 20 mM Tris pH 7.5, 50 mM NaCl, 1mM DTT, 1mM ADP, 1mM AlF3, 10 mM NaF, 2mM MgCl2, 0.5% glycerol, 0.005% Triton X-100 |
Grid | Model: Quantifoil R2/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: OTHER |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 279.15 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: 47.9 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |