+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-23908 | ||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | RNA polymerase II initially transcribing complex (ITC) | ||||||||||||||||||
![]() | ITC | ||||||||||||||||||
![]() |
| ||||||||||||||||||
![]() | PIC / TFIIH / transcription / ITC / RNA polymerase II | ||||||||||||||||||
Function / homology | ![]() : / regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / : / TFIIH-class transcription factor complex binding / RNA polymerase III preinitiation complex assembly ...: / regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / : / TFIIH-class transcription factor complex binding / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex / positive regulation of mitotic recombination / nucleotide-excision repair factor 3 complex / RNA polymerase I general transcription initiation factor binding / transcription factor TFIIE complex / nucleotide-excision repair, preincision complex assembly / DNA translocase activity / regulation of transcription by RNA polymerase III / hydrolase activity, acting on acid anhydrides, in phosphorus-containing anhydrides / TFIIF-class transcription factor complex binding / transcriptional start site selection at RNA polymerase II promoter / transcription factor TFIIF complex / RNA polymerase I preinitiation complex assembly / transcription factor TFIIA complex / transcription factor TFIIH core complex / transcription factor TFIIH holo complex / cyclin-dependent protein serine/threonine kinase activator activity / : / DNA binding, bending / transcription preinitiation complex / 3'-5' DNA helicase activity / Processing of Capped Intron-Containing Pre-mRNA / DNA 3'-5' helicase / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / poly(A)+ mRNA export from nucleus / mRNA Capping / RNA polymerase II transcribes snRNA genes / TP53 Regulates Transcription of DNA Repair Genes / 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 / RNA Polymerase II Pre-transcription Events / transcription factor TFIID complex / RNA polymerase II general transcription initiation factor activity / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Formation of TC-NER Pre-Incision Complex / RNA Polymerase I Promoter Escape / nucleolar large rRNA transcription by RNA polymerase I / Gap-filling DNA repair synthesis and ligation in TC-NER / RNA polymerase II complex binding / Estrogen-dependent gene expression / ATPase activator activity / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / protein phosphatase activator activity / positive regulation of translational initiation / Dual incision in TC-NER / RNA polymerase I complex / tRNA transcription by RNA polymerase III / RNA polymerase III complex / RNA polymerase II core promoter sequence-specific DNA binding / transcription-coupled nucleotide-excision repair / DNA helicase activity / RNA polymerase II, core complex / RNA polymerase II preinitiation complex assembly / translation initiation factor binding / TBP-class protein binding / DNA-directed RNA polymerase activity / isomerase activity / nucleotide-excision repair / positive regulation of transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / DNA-templated transcription initiation / transcription elongation by RNA polymerase II / P-body / ribonucleoside binding / : / : / : / DNA-directed RNA polymerase / disordered domain specific binding / single-stranded DNA binding / double-stranded DNA binding / DNA helicase / DNA-binding transcription factor binding / transcription regulator complex / RNA polymerase II-specific DNA-binding transcription factor binding / forked DNA-dependent helicase activity / single-stranded 3'-5' DNA helicase activity / four-way junction helicase activity / double-stranded DNA helicase activity / damaged DNA binding / transcription by RNA polymerase II / nucleic acid binding / single-stranded RNA binding / protein dimerization activity / chromatin extrusion motor activity / ATP-dependent H2AZ histone chaperone activity Similarity search - Function | ||||||||||||||||||
Biological species | ![]() ![]() | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||||||||||||||
![]() | Yang C / Fujiwara R | ||||||||||||||||||
Funding support | ![]()
| ||||||||||||||||||
![]() | ![]() Title: Structural visualization of de novo transcription initiation by Saccharomyces cerevisiae RNA polymerase II. Authors: Chun Yang / Rina Fujiwara / Hee Jong Kim / Pratik Basnet / Yunye Zhu / Jose J Gorbea Colón / Stefan Steimle / Benjamin A Garcia / Craig D Kaplan / Kenji Murakami / ![]() Abstract: Previous structural studies of the initiation-elongation transition of RNA polymerase II (pol II) transcription have relied on the use of synthetic oligonucleotides, often artificially discontinuous ...Previous structural studies of the initiation-elongation transition of RNA polymerase II (pol II) transcription have relied on the use of synthetic oligonucleotides, often artificially discontinuous to capture pol II in the initiating state. Here, we report multiple structures of initiation complexes converted de novo from a 33-subunit yeast pre-initiation complex (PIC) through catalytic activities and subsequently stalled at different template positions. We determine that PICs in the initially transcribing complex (ITC) can synthesize a transcript of ∼26 nucleotides before transitioning to an elongation complex (EC) as determined by the loss of general transcription factors (GTFs). Unexpectedly, transition to an EC was greatly accelerated when an ITC encountered a downstream EC stalled at promoter proximal regions and resulted in a collided head-to-end dimeric EC complex. Our structural analysis reveals a dynamic state of TFIIH, the largest of GTFs, in PIC/ITC with distinct functional consequences at multiple steps on the pathway to elongation. | ||||||||||||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 93 MB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 56.5 KB 56.5 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 9.1 KB | Display | ![]() |
Images | ![]() | 49.1 KB | ||
Filedesc metadata | ![]() | 14.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 373.3 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 372.9 KB | Display | |
Data in XML | ![]() | 4.3 KB | Display | |
Data in CIF | ![]() | 5 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7ml4MC ![]() 7meiC ![]() 7mk9C ![]() 7mkaC ![]() 7ml0C ![]() 7ml1C ![]() 7ml2C ![]() 7ml3C C: citing same article ( M: atomic model generated by this map |
---|---|
Similar structure data | |
EM raw data | ![]() Data size: 14.3 TB Data #1: raw micrographs for PIC + ITC maps [micrographs - multiframe] Data #2: raw micrographs for EC+EC map [micrographs - multiframe]) |
-
Links
EMDB pages | ![]() ![]() |
---|---|
Related items in Molecule of the Month |
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | ITC | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
+Entire : RNA polymerase II initially transcribing complex (ITC)
+Supramolecule #1: RNA polymerase II initially transcribing complex (ITC)
+Supramolecule #2: Pol II + TFIIB + RNA
+Supramolecule #3: DNA+TBP+TFIIA+TFIIE+TFIIF
+Supramolecule #4: TFIIH
+Macromolecule #1: Transcription initiation factor IIF subunit alpha
+Macromolecule #2: Transcription initiation factor IIF subunit beta
+Macromolecule #3: DNA-directed RNA polymerase II subunit RPB4
+Macromolecule #4: DNA-directed RNA polymerase II subunit RPB7
+Macromolecule #5: Transcription initiation factor IIB
+Macromolecule #6: DNA-directed RNA polymerase subunit
+Macromolecule #7: DNA-directed RNA polymerase subunit beta
+Macromolecule #8: DNA-directed RNA polymerase II subunit RPB3
+Macromolecule #9: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+Macromolecule #10: DNA-directed RNA polymerases I,II,and III subunit RPABC2
+Macromolecule #11: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+Macromolecule #12: DNA-directed RNA polymerase II subunit RPB9
+Macromolecule #13: DNA-directed RNA polymerases II subunit RPABC5
+Macromolecule #14: DNA-directed RNA polymerase II subunit RPB11
+Macromolecule #15: DNA-directed RNA polymerases II subunit RPABC4
+Macromolecule #16: BJ4_G0050160.mRNA.1.CDS.1
+Macromolecule #17: General transcription and DNA repair factor IIH helicase subunit XPD
+Macromolecule #18: General transcription and DNA repair factor IIH subunit TFB4
+Macromolecule #19: General transcription and DNA repair factor IIH
+Macromolecule #20: Tfb1
+Macromolecule #21: General transcription and DNA repair factor IIH helicase subunit XPB
+Macromolecule #22: General transcription and DNA repair factor IIH subunit TFB5
+Macromolecule #23: RNA polymerase II transcription factor B subunit 2
+Macromolecule #24: Transcription initiation factor IIE subunit beta
+Macromolecule #25: Transcription initiation factor IIA large subunit
+Macromolecule #26: Transcription initiation factor IIA subunit 2
+Macromolecule #29: TATA-box-binding protein
+Macromolecule #30: Transcription initiation factor IIE subunit alpha
+Macromolecule #27: non-template strand DNA
+Macromolecule #28: template strand DNA
+Macromolecule #31: RNA
+Macromolecule #32: ZINC ION
+Macromolecule #33: MAGNESIUM ION
+Macromolecule #34: IRON/SULFUR CLUSTER
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
![]() | single particle reconstruction |
Aggregation state | particle |
-
Sample preparation
Buffer | pH: 7.6 |
---|---|
Vitrification | Cryogen name: ETHANE |
-
Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 45.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |