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- EMDB-50732: RNA Polymerase III Class III Melting Pre-Initiation Complex (MC) -
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Basic information
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Title | RNA Polymerase III Class III Melting Pre-Initiation Complex (MC) | |||||||||
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![]() | RNA Polymerase III / SNAPc / TFIIIB / snRNA / Transcription | |||||||||
Function / homology | ![]() snRNA-activating protein complex / snRNA transcription / transcription preinitiation complex assembly / snRNA transcription by RNA polymerase III / bent DNA binding / RNA Polymerase III Chain Elongation / RNA polymerase III preinitiation complex assembly / TFIIIC-class transcription factor complex binding / RNA Polymerase III Transcription Termination / transcription factor TFIIIB complex ...snRNA-activating protein complex / snRNA transcription / transcription preinitiation complex assembly / snRNA transcription by RNA polymerase III / bent DNA binding / RNA Polymerase III Chain Elongation / RNA polymerase III preinitiation complex assembly / TFIIIC-class transcription factor complex binding / RNA Polymerase III Transcription Termination / transcription factor TFIIIB complex / calcitonin gene-related peptide receptor activity / DNA/RNA hybrid binding / RNA polymerase III type 3 promoter sequence-specific DNA binding / RNA polymerase transcription factor SL1 complex / regulation of transcription by RNA polymerase I / snRNA transcription by RNA polymerase II / regulation of transcription by RNA polymerase III / RPAP3/R2TP/prefoldin-like complex / RNA polymerase III general transcription initiation factor activity / RNA polymerase I core promoter sequence-specific DNA binding / DNA polymerase III complex / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Polymerase III Transcription Initiation From Type 3 Promoter / Cytosolic sensors of pathogen-associated DNA / transcription factor TFIIA complex / RNA Polymerase III Abortive And Retractive Initiation / female germ cell nucleus / male pronucleus / female pronucleus / positive regulation of innate immune response / RNA polymerase II general transcription initiation factor binding / nucleobase-containing compound metabolic process / Abortive elongation of HIV-1 transcript in the absence of Tat / FGFR2 alternative splicing / RNA Polymerase I Transcription Termination / MicroRNA (miRNA) biogenesis / transcription preinitiation complex / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / transcription factor TFIID complex / RNA polymerase II general transcription initiation factor activity / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / termination of RNA polymerase III transcription / mRNA Capping / HIV Transcription Initiation / RNA Polymerase II HIV Promoter Escape / Transcription of the HIV genome / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / transcription initiation at RNA polymerase III promoter / mRNA Splicing - Minor Pathway / PIWI-interacting RNA (piRNA) biogenesis / aryl hydrocarbon receptor binding / transcription by RNA polymerase I / TFIIB-class transcription factor binding / RNA Polymerase I Transcription Initiation / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / Processing of Capped Intron-Containing Pre-mRNA / transcription by RNA polymerase III / RNA polymerase II transcribes snRNA genes / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / positive regulation of transcription initiation by RNA polymerase II / neuropeptide signaling pathway / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / RNA polymerase I complex / tRNA transcription by RNA polymerase III / RNA polymerase III complex / transcription elongation by RNA polymerase I / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase II core promoter sequence-specific DNA binding / core promoter sequence-specific DNA binding / RNA polymerase II, core complex / : / RNA polymerase II preinitiation complex assembly / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / RNA Polymerase II Pre-transcription Events / TBP-class protein binding / DNA-directed RNA polymerase activity / Inhibition of DNA recombination at telomere / mRNA Splicing - Major Pathway / positive regulation of interferon-beta production / male germ cell nucleus / acrosomal vesicle / SIRT1 negatively regulates rRNA expression / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / DNA-templated transcription initiation / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / mRNA transcription by RNA polymerase II Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.51 Å | |||||||||
![]() | Shah SZ / Ramsay EP / Cecatiello V / Perry TN / Vannini A | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural insights into distinct mechanisms of RNA polymerase II and III recruitment to snRNA promoters. Authors: Syed Zawar Shah / Thomas N Perry / Andrea Graziadei / Valentina Cecatiello / Thangavelu Kaliyappan / Agata D Misiaszek / Christoph W Müller / Ewan P Ramsay / Alessandro Vannini / ![]() ![]() ![]() ![]() Abstract: RNA polymerase III (Pol III) transcribes short, essential RNAs, including the U6 small nuclear RNA (snRNA). At U6 snRNA genes, Pol III is recruited by the snRNA Activating Protein Complex (SNAPc) and ...RNA polymerase III (Pol III) transcribes short, essential RNAs, including the U6 small nuclear RNA (snRNA). At U6 snRNA genes, Pol III is recruited by the snRNA Activating Protein Complex (SNAPc) and a Brf2-containing TFIIIB complex, forming a pre-initiation complex (PIC). Uniquely, SNAPc also recruits Pol II at the remaining splicesosomal snRNA genes (U1, 2, 4 and 5). The mechanism of SNAPc cross-polymerase engagement and the role of the SNAPC2 and SNAPC5 subunits remain poorly defined. Here, we present cryo-EM structures of the full-length SNAPc-containing Pol III PIC assembled on the U6 snRNA promoter in the open and melting states at 3.2-4.2 Å resolution. The structural comparison revealed differences with the Saccharomyces cerevisiae Pol III PIC and the basis of selective SNAPc engagement within Pol III and Pol II PICs. Additionally, crosslinking mass spectrometry localizes SNAPC2 and SNAPC5 near the promoter DNA, expanding upon existing descriptions of snRNA Pol III PIC structure. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 16.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 54.5 KB 54.5 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.9 KB | Display | ![]() |
Images | ![]() | 47.9 KB | ||
Filedesc metadata | ![]() | 14.2 KB | ||
Others | ![]() ![]() | 262.1 MB 262.1 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1 MB | Display | |
Data in XML | ![]() | 22.2 KB | Display | |
Data in CIF | ![]() | 28.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9fsqMC ![]() 9fsoC ![]() 9fspC ![]() 9fsrC ![]() 9fssC 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: 0.955 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #2
File | emd_50732_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_50732_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
+Entire : RNA Polymerase III Class III Melting Pre-Initiation Complex
+Supramolecule #1: RNA Polymerase III Class III Melting Pre-Initiation Complex
+Supramolecule #2: SNAPc
+Supramolecule #3: TFIIIB
+Supramolecule #4: DNA
+Supramolecule #5: Human RNA Polymerase III
+Macromolecule #1: DNA-directed RNA polymerase III subunit RPC1
+Macromolecule #2: DNA-directed RNA polymerase III subunit RPC2
+Macromolecule #3: DNA-directed RNA polymerase III subunit RPC3
+Macromolecule #4: DNA-directed RNA polymerase III subunit RPC4
+Macromolecule #5: DNA-directed RNA polymerase III subunit RPC5
+Macromolecule #6: DNA-directed RNA polymerase III subunit RPC6
+Macromolecule #7: DNA-directed RNA polymerase III subunit RPC7
+Macromolecule #8: DNA-directed RNA polymerase III subunit RPC8
+Macromolecule #9: DNA-directed RNA polymerase III subunit RPC9
+Macromolecule #10: DNA-directed RNA polymerase III subunit RPC10
+Macromolecule #11: DNA-directed RNA polymerases I and III subunit RPAC1
+Macromolecule #12: DNA-directed RNA polymerases I and III subunit RPAC2
+Macromolecule #13: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+Macromolecule #14: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+Macromolecule #15: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+Macromolecule #16: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+Macromolecule #17: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+Macromolecule #18: TATA-box-binding protein
+Macromolecule #19: Transcription factor IIIB 50 kDa subunit
+Macromolecule #20: Transcription factor TFIIIB component B'' homolog
+Macromolecule #21: snRNA-activating protein complex subunit 1
+Macromolecule #22: snRNA-activating protein complex subunit 4
+Macromolecule #25: snRNA-activating protein complex subunit 3
+Macromolecule #23: U6_2_Template
+Macromolecule #24: U6_2_Non template
+Macromolecule #26: ZINC ION
+Macromolecule #27: MAGNESIUM ION
+Macromolecule #28: IRON/SULFUR CLUSTER
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 150 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. | ||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: TFS Selectris X / Energy filter - Slit width: 10 eV |
Details | FEI Titan Krios G4 (operating at 300 KeV) |
Image recording | Film or detector model: TFS FALCON 4i (4k x 4k) / Number real images: 17291 / Average exposure time: 5.3 sec. / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.75 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |