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- EMDB-14383: Mammalian Dicer in the "pre-dicing state" with pre-miR-15a substrate -
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Open data
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Basic information
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Title | Mammalian Dicer in the "pre-dicing state" with pre-miR-15a substrate | |||||||||
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![]() | endoribonuclease / dsRNA / complex / gene silencing / post-transcriptional / catalytic complex / cytoplasm / RNA BINDING PROTEIN | |||||||||
Function / homology | ![]() regulation of muscle cell apoptotic process / myoblast differentiation involved in skeletal muscle regeneration / MicroRNA (miRNA) biogenesis / Small interfering RNA (siRNA) biogenesis / ganglion development / hair follicle cell proliferation / zygote asymmetric cell division / regulation of oligodendrocyte differentiation / cardiac neural crest cell development involved in outflow tract morphogenesis / positive regulation of endothelial cell-matrix adhesion via fibronectin ...regulation of muscle cell apoptotic process / myoblast differentiation involved in skeletal muscle regeneration / MicroRNA (miRNA) biogenesis / Small interfering RNA (siRNA) biogenesis / ganglion development / hair follicle cell proliferation / zygote asymmetric cell division / regulation of oligodendrocyte differentiation / cardiac neural crest cell development involved in outflow tract morphogenesis / positive regulation of endothelial cell-matrix adhesion via fibronectin / olfactory bulb interneuron differentiation / regulation of enamel mineralization / positive regulation of establishment of endothelial barrier / positive regulation of hepatic stellate cell proliferation / trophectodermal cell proliferation / regulation of miRNA metabolic process / regulation of odontogenesis of dentin-containing tooth / spermatogonial cell division / regulation of RNA metabolic process / peripheral nervous system myelin formation / regulation of epithelial cell differentiation / regulation of regulatory T cell differentiation / regulation of Notch signaling pathway / spinal cord motor neuron differentiation / global gene silencing by mRNA cleavage / epidermis morphogenesis / negative regulation of Schwann cell proliferation / reproductive structure development / ribonuclease III / positive regulation of myelination / apoptotic DNA fragmentation / inner ear receptor cell development / nerve development / meiotic spindle organization / positive regulation of Schwann cell differentiation / RISC-loading complex / deoxyribonuclease I activity / RISC complex assembly / regulatory ncRNA-mediated post-transcriptional gene silencing / intestinal epithelial cell development / ribonuclease III activity / pericentric heterochromatin formation / miRNA processing / regulation of stem cell differentiation / pre-miRNA processing / regulation of viral genome replication / siRNA processing / mRNA stabilization / digestive tract development / embryonic hindlimb morphogenesis / cartilage development / cardiac muscle cell development / RISC complex / embryonic limb morphogenesis / miRNA binding / positive regulation of vascular endothelial cell proliferation / regulation of myelination / regulation of neuron differentiation / negative regulation of glial cell proliferation / hair follicle morphogenesis / stem cell population maintenance / branching morphogenesis of an epithelial tube / positive regulation of miRNA metabolic process / endoplasmic reticulum-Golgi intermediate compartment / regulation of neurogenesis / hair follicle development / positive regulation of collagen biosynthetic process / postsynaptic density, intracellular component / spindle assembly / RNA processing / spleen development / positive regulation of endothelial cell migration / helicase activity / neuron projection morphogenesis / post-embryonic development / regulation of protein phosphorylation / lung development / multicellular organism growth / cerebral cortex development / rRNA processing / gene expression / growth cone / regulation of inflammatory response / regulation of gene expression / angiogenesis / defense response to virus / cell population proliferation / regulation of cell cycle / axon / glutamatergic synapse / dendrite / positive regulation of gene expression / perinuclear region of cytoplasm / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / DNA binding / RNA binding / ATP binding / nucleus / metal ion binding Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.19 Å | |||||||||
![]() | Zanova M / Zapletal D / Kubicek K / Stefl R / Pinkas M / Novacek J | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural and functional basis of mammalian microRNA biogenesis by Dicer. Authors: David Zapletal / Eliska Taborska / Josef Pasulka / Radek Malik / Karel Kubicek / Martina Zanova / Christian Much / Marek Sebesta / Valeria Buccheri / Filip Horvat / Irena Jenickova / ...Authors: David Zapletal / Eliska Taborska / Josef Pasulka / Radek Malik / Karel Kubicek / Martina Zanova / Christian Much / Marek Sebesta / Valeria Buccheri / Filip Horvat / Irena Jenickova / Michaela Prochazkova / Jan Prochazka / Matyas Pinkas / Jiri Novacek / Diego F Joseph / Radislav Sedlacek / Carrie Bernecky / Dónal O'Carroll / Richard Stefl / Petr Svoboda / ![]() ![]() ![]() ![]() ![]() Abstract: MicroRNA (miRNA) and RNA interference (RNAi) pathways rely on small RNAs produced by Dicer endonucleases. Mammalian Dicer primarily supports the essential gene-regulating miRNA pathway, but how it is ...MicroRNA (miRNA) and RNA interference (RNAi) pathways rely on small RNAs produced by Dicer endonucleases. Mammalian Dicer primarily supports the essential gene-regulating miRNA pathway, but how it is specifically adapted to miRNA biogenesis is unknown. We show that the adaptation entails a unique structural role of Dicer's DExD/H helicase domain. Although mice tolerate loss of its putative ATPase function, the complete absence of the domain is lethal because it assures high-fidelity miRNA biogenesis. Structures of murine Dicer•-miRNA precursor complexes revealed that the DExD/H domain has a helicase-unrelated structural function. It locks Dicer in a closed state, which facilitates miRNA precursor selection. Transition to a cleavage-competent open state is stimulated by Dicer-binding protein TARBP2. Absence of the DExD/H domain or its mutations unlocks the closed state, reduces substrate selectivity, and activates RNAi. Thus, the DExD/H domain structurally contributes to mammalian miRNA biogenesis and underlies mechanistical partitioning of miRNA and RNAi pathways. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 196.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 21.8 KB 21.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.4 KB | Display | ![]() |
Images | ![]() | 84.4 KB | ||
Filedesc metadata | ![]() | 7.7 KB | ||
Others | ![]() ![]() | 200.7 MB 200.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1 MB | Display | |
Data in XML | ![]() | 21.8 KB | Display | |
Data in CIF | ![]() | 28.2 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7yymMC ![]() 7yynC ![]() 7yz4C ![]() 7zpiC ![]() 7zpjC ![]() 7zpkC 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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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.828 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: half map B
File | emd_14383_half_map_1.map | ||||||||||||
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Annotation | half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map A
File | emd_14383_half_map_2.map | ||||||||||||
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Annotation | half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Pre-dicing state of mouse somatic dicer with pre-mir-15a
Entire | Name: Pre-dicing state of mouse somatic dicer with pre-mir-15a |
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Components |
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-Supramolecule #1: Pre-dicing state of mouse somatic dicer with pre-mir-15a
Supramolecule | Name: Pre-dicing state of mouse somatic dicer with pre-mir-15a type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Supramolecule #2: Pre-dicing state of mouse somatic dicer
Supramolecule | Name: Pre-dicing state of mouse somatic dicer / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
-Supramolecule #3: pre-mir-15a
Supramolecule | Name: pre-mir-15a / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2 |
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Source (natural) | Organism: synthetic construct (others) |
-Macromolecule #1: Endoribonuclease Dicer
Macromolecule | Name: Endoribonuclease Dicer / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: ribonuclease III |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 226.809594 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MVWSHPQFEK GGGSGGGSGG SAWSHPQFEK GDYPYDVPDY AGTENLYFQG LVDMKSPALQ PLSMAGLQLM TPASSPMGPF FGLPWQQEA IHDNIYTPRK YQVELLEAAL DHNTIVCLNT GSGKTFIAVL LTKELAHQIR GDLNPHAKRT VFLVNSANQV A QQVSAVRT ...String: MVWSHPQFEK GGGSGGGSGG SAWSHPQFEK GDYPYDVPDY AGTENLYFQG LVDMKSPALQ PLSMAGLQLM TPASSPMGPF FGLPWQQEA IHDNIYTPRK YQVELLEAAL DHNTIVCLNT GSGKTFIAVL LTKELAHQIR GDLNPHAKRT VFLVNSANQV A QQVSAVRT HSDLKVGEYS DLEVNASWTK ERWSQEFTKH QVLIMTCYVA LTVLKNGYLS LSDINLLVFD ECHLAILDHP YR EIMKLCE SCPSCPRILG LTASILNGKC DPEELEEKIQ KLERILRSDA ETATDLVVLD RYTSQPCEIV VDCGPFTDRS GLY ERLLME LEAALDFIND CNVAVHSKER DSTLISKQIL SDCRAVLVVL GPWCADKVAG MMVRELQKYI KHEQEELHRK FLLF TDTLL RKIHALCEEY FSPASLDLKY VTPKVMKLLE ILRKYKPYER QQFESVEWYN NRNQDNYVSW SDSEDDDDDE EIEEK EKPE TNFPSPFTNI LCGIIFVERR YTAVVLNRLI KEAGKQDPEL AYISSNFITG HGIGKNQPRS KQMEAEFRKQ EEVLRK FRA HETNLLIATS VVEEGVDIPK CNLVVRFDLP TEYRSYVQSK GRARAPISNY VMLADTDKIK SFEEDLKTYK AIEKILR NK CSKSADGAEA DVHAGVDDED AFPPYVLRPD DGGPRVTINT AIGHINRYCA RLPSDPFTHL APKCRTRELP DGTFYSTL Y LPINSPLRAS IVGPPMDSVR LAERVVALIC CEKLHKIGEL DEHLMPVGKE TVKYEEELDL HDEEETSVPG RPGSTKRRQ CYPKAIPECL RESYPKPDQP CYLYVIGMVL TTPLPDELNF RRRKLYPPED TTRCFGILTA KPIPQIPHFP VYTRSGEVTI SIELKKSGF TLSQQMLELI TRLHQYIFSH ILRLEKPALE FKPTGAESAY CVLPLNVVND SGTLDIDFKF MEDIEKSEAR I GIPSTKYS KETPFVFKLE DYQDAVIIPR YRNFDQPHRF YVADVYTDLT PLSKFPSPEY ETFAEYYKTK YNLDLTNLNQ PL LDVDHTS SRLNLLTPRH LNQKGKALPL SSAEKRKAKW ESLQNKQILV PELCAIHPIP ASLWRKAVCL PSILYRLHCL LTA EELRAQ TASDAGVGVR SLPVDFRYPN LDFGWKKSID SKSFISSCNS SLAESDNYCK HSTTVVPEHA AHQGATRPSL ENHD QMSVN CKRLPAESPA KLQSEVSTDL TAINGLSYNK NLANGSYDLV NRDFCQGNQL NYFKQEIPVQ PTTSYPIQNL YNYEN QPKP SNECPLLSNT YLDGNANTST SDGSPAVSTM PAMMNAVKAL KDRMDSEQSP SVGYSSRTLG PNPGLILQAL TLSNAS DGF NLERLEMLGD SFLKHAITTY LFCTYPDAHE GRLSYMRSKK VSNCNLYRLG KKKGLPSRMV VSIFDPPVNW LPPGYVV NQ DKSNSEKWEK DEMTKDCLLA NGKLGEACEE EEDLTWRAPK EEAEDEDDFL EYDQEHIQFI DSMLMGSGAF VRKISLSP F SASDSAYEWK MPKKASLGSM PFASGLEDFD YSSWDAMCYL DPSKAVEEDD FVVGFWNPSE ENCGVDTGKQ SISYDLHTE QCIADKSIAD CVAALLGCYL TSCGERAAQL FLCSLGLKVL PVIKRTSREK ALDPAQENGS SQQKSLSGSC ASPVGPRSSA GKDLEYGCL KIPPRCMFDH PDAEKTLNHL ISGFETFEKK INYRFKNKAY LLQAFTHASY HYNTITDCYQ RLEFLGDAIL D YLITKHLY EDPRQHSPGV LTDLRSALVN NTIFASLAVK YDYHKYFKAV SPELFHVIDD FVKFQLEKNE MQGMDSELRR SE EDEEKEE DIEVPKAMGD IFASLAGAIY MDSGMSLEVV WQVYYPMMQP LIEKFSANVP RSPVRELLEM EPETAKFSPA ERT YDGKVR VTVEVVGKGK FKGVGRSYRI AKSAAARRAL RSLKANQPQV PNSGRGENLY FQGASDYKDH DGDYKDHDGS HHHH HHHH UniProtKB: Endoribonuclease Dicer |
-Macromolecule #2: 59-nt precursor of miR-15a
Macromolecule | Name: 59-nt precursor of miR-15a / type: rna / ID: 2 / Number of copies: 1 |
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Source (natural) | Organism: synthetic construct (others) |
Molecular weight | Theoretical: 19.017279 KDa |
Sequence | String: UAGCAGCACA UAAUGGUUUG UGGAUGUUGA AAAGGUGCAG GCCAUACUGU GCUGCCUCA |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.20 mg/mL | ||||||||||
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Buffer | pH: 8 Component:
Details: The buffer was always prepared fresh in RNAse-free manner. | ||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV / Details: Described in STAR methods. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 16601 / Average electron dose: 55.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Details | Endoribonuclease mouse Dicer from AlphaFold database was used as initial model. Initial local fitting was done using Chimera and then Coot's Real Space Refine Zone. |
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Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Overall B value: 134.8 Target criteria: Ramachandran Plot, Rotamer Analysis, Density Analysis |
Output model | ![]() PDB-7yym: |
-Atomic model buiding 2
Details | PHENIX Real-space refinement was used for flexible fitting. |
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Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Overall B value: 134.8 / Target criteria: Correlation coefficient |
Output model | ![]() PDB-7yym: |
-Atomic model buiding 3
Details | ISOLDE was used for flexible fitting with torsion restraints defined for polypeptide and distance restraints for polyribonucleotide chain. |
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Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Overall B value: 134.8 Target criteria: Ramachandran Plot, Rotamer Analysis, Clash Score |
Output model | ![]() PDB-7yym: |