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- PDB-9fsq: RNA Polymerase III Class III Melting Pre-Initiation Complex (MC) -

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Basic information

Entry
Database: PDB / ID: 9fsq
TitleRNA Polymerase III Class III Melting Pre-Initiation Complex (MC)
Components
  • (DNA-directed RNA polymerase III subunit ...) x 10
  • (DNA-directed RNA polymerases I and III subunit ...) x 2
  • (DNA-directed RNA polymerases I, II, and III subunit ...) x 5
  • (Transcription factor ...) x 2
  • (snRNA-activating protein complex subunit ...) x 3
  • TATA-box-binding protein
  • U6_2_Non template
  • U6_2_Template
KeywordsTRANSCRIPTION / RNA Polymerase III / SNAPc / TFIIIB / snRNA
Function / homology
Function and homology information


snRNA-activating protein complex / transcription preinitiation complex assembly / snRNA transcription / snRNA transcription by RNA polymerase III / bent DNA binding / RNA Polymerase III Chain Elongation / calcitonin gene-related peptide receptor activity / RNA Polymerase III Transcription Termination / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex ...snRNA-activating protein complex / transcription preinitiation complex assembly / snRNA transcription / snRNA transcription by RNA polymerase III / bent DNA binding / RNA Polymerase III Chain Elongation / calcitonin gene-related peptide receptor activity / RNA Polymerase III Transcription Termination / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex / TFIIIC-class transcription factor complex binding / DNA/RNA hybrid binding / RNA polymerase III type 3 promoter sequence-specific DNA binding / regulation of transcription by RNA polymerase I / RNA polymerase transcription factor SL1 complex / regulation of transcription by RNA polymerase III / snRNA transcription by RNA polymerase II / 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 / Cytosolic sensors of pathogen-associated DNA / 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 / RNA Polymerase III Abortive And Retractive Initiation / transcription factor TFIIA complex / female germ cell nucleus / positive regulation of innate immune response / male pronucleus / female pronucleus / 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 / transcription preinitiation complex / MicroRNA (miRNA) biogenesis / 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 / PIWI-interacting RNA (piRNA) biogenesis / 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 / aryl hydrocarbon receptor binding / transcription by RNA polymerase I / RNA Polymerase I Transcription Initiation / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / TFIIB-class transcription factor binding / Processing of Capped Intron-Containing Pre-mRNA / transcription by RNA polymerase III / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / RNA polymerase II transcribes snRNA genes / : / neuropeptide signaling pathway / positive regulation of transcription initiation by RNA polymerase II / RNA polymerase II core promoter sequence-specific DNA binding / transcription elongation by RNA polymerase I / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / tRNA transcription by RNA polymerase III / RNA polymerase I complex / RNA polymerase III complex / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase II, core complex / core promoter sequence-specific DNA binding / RNA Polymerase II Transcription Elongation / RNA polymerase II preinitiation complex assembly / Formation of RNA Pol II elongation complex / RNA Polymerase II Pre-transcription Events / TBP-class protein binding / positive regulation of interferon-beta production / Inhibition of DNA recombination at telomere / mRNA Splicing - Major Pathway / acrosomal vesicle / SIRT1 negatively regulates rRNA expression / male germ cell nucleus / 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
: / BRF2-like, C-terminal cyclin repeat / snRNA-activating protein complex subunit 1 / snRNA-activating protein complex, subunit 3 / : / Small nuclear RNA activating complex (SNAPc), subunit 1 / snRNA-activating protein complex (SNAPc), subunit 3 / Transcription factor TFIIIB component B'', Myb domain / Myb DNA-binding like / DNA-directed RNA polymerase III subunit RPC5, C-terminal ...: / BRF2-like, C-terminal cyclin repeat / snRNA-activating protein complex subunit 1 / snRNA-activating protein complex, subunit 3 / : / Small nuclear RNA activating complex (SNAPc), subunit 1 / snRNA-activating protein complex (SNAPc), subunit 3 / Transcription factor TFIIIB component B'', Myb domain / Myb DNA-binding like / DNA-directed RNA polymerase III subunit RPC5, C-terminal / DNA-directed RNA polymerase III subunit RPC5 C-terminal / DNA-directed RNA polymerase III subunit RPC4 / RNA polymerase III RPC4 / DNA-directed RNA polymerase III, subunit Rpc31 / DNA-directed RNA polymerase III subunit Rpc31 / Pol III subunit C11, C-terminal zinc ribbon / DNA-directed RNA polymerase III subunit Rpc5 / DNA-directed RNA polymerase III subunit RPC1, N-terminal / DNA-directed RNA polymerase III subunit RPC1, C-terminal / RPC5 protein / RNA polymerase III, subunit Rpc25 / DNA-directed RNA polymerase III subunit RPC9 / RNA polymerase III subunit Rpc25 / RNA polymerase Rpc34 / RNA polymerase III Rpc82, C -terminal / RNA polymerase Rpc34-like / DNA-directed RNA polymerase III subunit RPC3 / : / RNA polymerase Rpc34 subunit / RNA polymerase III subunit RPC82 / DNA-directed RNA polymerase III subunit RPC3, helical hairpin domain / POLR3C, C-terminal winged-helix domain / Myb-like domain profile. / RNA polymerase III subunit RPC82-related, helix-turn-helix / RNA polymerase III subunit RPC82 helix-turn-helix domain / DNA-directed RNA polymerase, subunit E/RPC8 / Myb-like DNA-binding domain / DNA-directed RNA polymerases I and III subunit AC19 / DNA-directed RNA polymerases I and III subunit AC40 / Zinc finger TFIIB-type profile. / Transcription factor TFIIB / Zinc finger, TFIIB-type / TFIIB zinc-binding / Myb-type HTH DNA-binding domain profile. / TATA-box binding protein, eukaryotic / TATA-box binding protein / TATA-box binding protein, conserved site / Transcription factor TFIID (or TATA-binding protein, TBP) / Transcription factor TFIID repeat signature. / SANT domain / Myb domain / TBP domain superfamily / RNA polymerase Rpb4/RPC9, core / DNA-directed RNA-polymerase II subunit / Rpb4/RPC9 superfamily / RNA polymerase subunit Rpb4/RPC9 / RNA polymerase Rpb4 / SANT SWI3, ADA2, N-CoR and TFIIIB'' DNA-binding domains / Cyclin-like superfamily / SANT/Myb domain / Zinc finger TFIIS-type signature. / HRDC-like superfamily / RNA polymerase Rpb7-like , N-terminal / RNA polymerase Rpb7-like, N-terminal domain superfamily / RNA polymerase subunit Rpb7-like / SHS2 domain found in N terminus of Rpb7p/Rpc25p/MJ0397 / RNA polymerase Rpb2, domain 5 / RNA polymerase Rpb2, domain 5 / RNA polymerase Rpb2, domain 4 / RNA polymerase Rpb2, domain 4 / DNA-directed RNA polymerase, M/15kDa subunit / RNA polymerases M/15 Kd subunit / RNA polymerase subunit 9 / DNA-directed RNA polymerase M, 15kDa subunit, conserved site / RNA polymerases M / 15 Kd subunits signature. / DNA-directed RNA polymerase subunit/transcription factor S / : / RNA polymerase, Rpb8 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / RNA polymerase Rpb8 / RNA polymerase subunit 8 / RNA polymerase, Rpb5, N-terminal / RNA polymerase Rpb5, N-terminal domain superfamily / RNA polymerase Rpb5, N-terminal domain / DNA-directed RNA polymerase, subunit RPB6 / DNA-directed RNA polymerase subunit RPABC5/Rpb10 / RNA polymerases, subunit N, zinc binding site / RNA polymerase subunit RPB10 / RNA polymerases N / 8 kDa subunit / RNA polymerases N / 8 Kd subunits signature. / DNA directed RNA polymerase, 7 kDa subunit / RNA polymerase archaeal subunit P/eukaryotic subunit RPABC4 / RNA polymerase, subunit H/Rpb5, conserved site / RNA polymerases H / 23 Kd subunits signature. / RNA polymerase subunit CX / DNA-directed RNA polymerase, 30-40kDa subunit, conserved site / DNA-directed RNA polymerase subunit Rpo3/Rpb3/RPAC1 / RNA polymerases D / 30 to 40 Kd subunits signature. / DNA-directed RNA polymerase Rpb11, 13-16kDa subunit, conserved site / DNA-directed RNA polymerase subunit Rpo11
Similarity search - Domain/homology
IRON/SULFUR CLUSTER / DNA / DNA (> 10) / Transcription factor TFIIIB component B'' homolog / DNA-directed RNA polymerase III subunit RPC1 / DNA-directed RNA polymerases I and III subunit RPAC1 / DNA-directed RNA polymerase III subunit RPC7 / DNA-directed RNA polymerase III subunit RPC9 / DNA-directed RNA polymerase III subunit RPC4 / DNA-directed RNA polymerases I and III subunit RPAC2 ...IRON/SULFUR CLUSTER / DNA / DNA (> 10) / Transcription factor TFIIIB component B'' homolog / DNA-directed RNA polymerase III subunit RPC1 / DNA-directed RNA polymerases I and III subunit RPAC1 / DNA-directed RNA polymerase III subunit RPC7 / DNA-directed RNA polymerase III subunit RPC9 / DNA-directed RNA polymerase III subunit RPC4 / DNA-directed RNA polymerases I and III subunit RPAC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / TATA-box-binding protein / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / snRNA-activating protein complex subunit 1 / snRNA-activating protein complex subunit 4 / snRNA-activating protein complex subunit 3 / DNA-directed RNA polymerase III subunit RPC3 / DNA-directed RNA polymerase III subunit RPC6 / Transcription factor IIIB 50 kDa subunit / DNA-directed RNA polymerase III subunit RPC5 / DNA-directed RNA polymerase III subunit RPC2 / DNA-directed RNA polymerase III subunit RPC10 / DNA-directed RNA polymerase III subunit RPC8
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.51 Å
AuthorsShah, S.Z. / Ramsay, E.P. / Cecatiello, V. / Perry, T.N. / Vannini, A.
Funding support Italy, 1items
OrganizationGrant numberCountry
Other government Italy
CitationJournal: Nat Commun / Year: 2025
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
DepositionJun 21, 2024Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jan 15, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: DNA-directed RNA polymerase III subunit RPC1
B: DNA-directed RNA polymerase III subunit RPC2
C: DNA-directed RNA polymerase III subunit RPC3
D: DNA-directed RNA polymerase III subunit RPC4
E: DNA-directed RNA polymerase III subunit RPC5
F: DNA-directed RNA polymerase III subunit RPC6
G: DNA-directed RNA polymerase III subunit RPC7
H: DNA-directed RNA polymerase III subunit RPC8
I: DNA-directed RNA polymerase III subunit RPC9
J: DNA-directed RNA polymerase III subunit RPC10
K: DNA-directed RNA polymerases I and III subunit RPAC1
L: DNA-directed RNA polymerases I and III subunit RPAC2
M: DNA-directed RNA polymerases I, II, and III subunit RPABC1
N: DNA-directed RNA polymerases I, II, and III subunit RPABC2
O: DNA-directed RNA polymerases I, II, and III subunit RPABC3
P: DNA-directed RNA polymerases I, II, and III subunit RPABC4
Q: DNA-directed RNA polymerases I, II, and III subunit RPABC5
R: TATA-box-binding protein
S: Transcription factor IIIB 50 kDa subunit
T: Transcription factor TFIIIB component B'' homolog
U: snRNA-activating protein complex subunit 1
W: snRNA-activating protein complex subunit 4
X: U6_2_Template
Y: U6_2_Non template
Z: snRNA-activating protein complex subunit 3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)1,149,06236
Polymers1,148,09825
Non-polymers96511
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1

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Components

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DNA-directed RNA polymerase III subunit ... , 10 types, 10 molecules ABCDEFGHIJ

#1: Protein DNA-directed RNA polymerase III subunit RPC1 / RNA polymerase III subunit C1 / DNA-directed RNA polymerase III largest subunit / DNA-directed RNA ...RNA polymerase III subunit C1 / DNA-directed RNA polymerase III largest subunit / DNA-directed RNA polymerase III subunit A / RNA polymerase III 155 kDa subunit / RPC155 / RNA polymerase III subunit C160


Mass: 155860.125 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O14802, DNA-directed RNA polymerase
#2: Protein DNA-directed RNA polymerase III subunit RPC2 / RNA polymerase III subunit C2 / C128 / DNA-directed RNA polymerase III 127.6 kDa polypeptide / DNA- ...RNA polymerase III subunit C2 / C128 / DNA-directed RNA polymerase III 127.6 kDa polypeptide / DNA-directed RNA polymerase III subunit B


Mass: 127953.891 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9NW08, DNA-directed RNA polymerase
#3: Protein DNA-directed RNA polymerase III subunit RPC3 / RNA polymerase III subunit C3 / DNA-directed RNA polymerase III subunit C / RNA polymerase III 62 ...RNA polymerase III subunit C3 / DNA-directed RNA polymerase III subunit C / RNA polymerase III 62 kDa subunit / RPC62


Mass: 60692.555 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BUI4
#4: Protein DNA-directed RNA polymerase III subunit RPC4 / RNA polymerase III subunit C4 / DNA-directed RNA polymerase III subunit D / Protein BN51 / RNA ...RNA polymerase III subunit C4 / DNA-directed RNA polymerase III subunit D / Protein BN51 / RNA polymerase III 47 kDa subunit / RPC53 homolog


Mass: 44471.875 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P05423
#5: Protein DNA-directed RNA polymerase III subunit RPC5 / RNA polymerase III subunit C5 / DNA-directed RNA polymerase III 80 kDa polypeptide


Mass: 80004.031 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9NVU0
#6: Protein DNA-directed RNA polymerase III subunit RPC6 / RNA polymerase III subunit C6 / DNA-directed RNA polymerase III subunit F / RNA polymerase III 39 ...RNA polymerase III subunit C6 / DNA-directed RNA polymerase III subunit F / RNA polymerase III 39 kDa subunit / RPC39


Mass: 35726.914 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9H1D9
#7: Protein DNA-directed RNA polymerase III subunit RPC7 / RNA polymerase III subunit C7 / DNA-directed RNA polymerase III subunit G / RNA polymerase III 32 ...RNA polymerase III subunit C7 / DNA-directed RNA polymerase III subunit G / RNA polymerase III 32 kDa apha subunit / RPC32-alpha / RNA polymerase III 32 kDa subunit / RPC32


Mass: 25953.510 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O15318
#8: Protein DNA-directed RNA polymerase III subunit RPC8 / RNA polymerase III subunit C8 / DNA-directed RNA polymerase III subunit H / RNA polymerase III ...RNA polymerase III subunit C8 / DNA-directed RNA polymerase III subunit H / RNA polymerase III subunit 22.9 kDa subunit / RPC22.9


Mass: 22938.846 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y535
#9: Protein DNA-directed RNA polymerase III subunit RPC9 / RNA polymerase III subunit C9 / Calcitonin gene-related peptide-receptor component protein / HsC17


Mass: 16893.990 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75575
#10: Protein DNA-directed RNA polymerase III subunit RPC10 / RNA polymerase III subunit C10 / DNA-directed RNA polymerase III subunit K / RNA polymerase III 12. ...RNA polymerase III subunit C10 / DNA-directed RNA polymerase III subunit K / RNA polymerase III 12.5 kDa subunit / RPC12.5 / RNA polymerase III subunit C11 / hRPC11


Mass: 12354.104 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y2Y1

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DNA-directed RNA polymerases I and III subunit ... , 2 types, 2 molecules KL

#11: Protein DNA-directed RNA polymerases I and III subunit RPAC1 / RNA polymerases I and III subunit AC1 / AC40 / DNA-directed RNA polymerases I and III 40 kDa ...RNA polymerases I and III subunit AC1 / AC40 / DNA-directed RNA polymerases I and III 40 kDa polypeptide / RPA40 / RPA39 / RPC40


Mass: 39301.672 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O15160
#12: Protein DNA-directed RNA polymerases I and III subunit RPAC2 / RNA polymerases I and III subunit AC2 / AC19 / DNA-directed RNA polymerase I subunit D / RNA ...RNA polymerases I and III subunit AC2 / AC19 / DNA-directed RNA polymerase I subunit D / RNA polymerase I 16 kDa subunit / RPA16 / RPC16 / hRPA19


Mass: 15259.222 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P0DPB6

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DNA-directed RNA polymerases I, II, and III subunit ... , 5 types, 5 molecules MNOPQ

#13: Protein DNA-directed RNA polymerases I, II, and III subunit RPABC1 / RNA polymerases I / II / and III subunit ABC1 / DNA-directed RNA polymerase II 23 kDa polypeptide / ...RNA polymerases I / II / and III subunit ABC1 / DNA-directed RNA polymerase II 23 kDa polypeptide / DNA-directed RNA polymerase II subunit E / RPB5 homolog / XAP4


Mass: 24584.223 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P19388
#14: Protein DNA-directed RNA polymerases I, II, and III subunit RPABC2 / RNA polymerases I / II / and III subunit ABC2 / DNA-directed RNA polymerase II subunit F / DNA- ...RNA polymerases I / II / and III subunit ABC2 / DNA-directed RNA polymerase II subunit F / DNA-directed RNA polymerases I / and III 14.4 kDa polypeptide / RPABC14.4 / RPB14.4 / RPB6 homolog / RPC15


Mass: 14491.026 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P61218
#15: Protein DNA-directed RNA polymerases I, II, and III subunit RPABC3 / RNA polymerases I / II / and III subunit ABC3 / DNA-directed RNA polymerase II subunit H / DNA- ...RNA polymerases I / II / and III subunit ABC3 / DNA-directed RNA polymerase II subunit H / DNA-directed RNA polymerases I / and III 17.1 kDa polypeptide / RPB17 / RPB8 homolog / hRPB8


Mass: 17162.273 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P52434
#16: Protein DNA-directed RNA polymerases I, II, and III subunit RPABC4 / RNA polymerases I / II / and III subunit ABC4 / ABC10-alpha / DNA-directed RNA polymerase II ...RNA polymerases I / II / and III subunit ABC4 / ABC10-alpha / DNA-directed RNA polymerase II subunit K / RNA polymerase II 7.0 kDa subunit / RPB7.0 / RPB10alpha


Mass: 7018.244 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P53803
#17: Protein DNA-directed RNA polymerases I, II, and III subunit RPABC5 / RNA polymerases I / II / and III subunit ABC5 / DNA-directed RNA polymerase III subunit L / RNA ...RNA polymerases I / II / and III subunit ABC5 / DNA-directed RNA polymerase III subunit L / RNA polymerase II 7.6 kDa subunit / RPB7.6 / RPB10 homolog


Mass: 7655.123 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62875

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Protein , 1 types, 1 molecules R

#18: Protein TATA-box-binding protein / TATA sequence-binding protein / TATA-binding factor / TATA-box factor / Transcription initiation ...TATA sequence-binding protein / TATA-binding factor / TATA-box factor / Transcription initiation factor TFIID TBP subunit


Mass: 22615.730 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: TBP, GTF2D1, TF2D, TFIID / Production host: Escherichia coli (E. coli) / References: UniProt: P20226

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Transcription factor ... , 2 types, 2 molecules ST

#19: Protein Transcription factor IIIB 50 kDa subunit / TFIIIB50 / hTFIIIB50 / B-related factor 2 / BRF-2 / hBRFU


Mass: 46587.355 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: BRF2, BRFU, PRO1470 / Production host: Escherichia coli (E. coli) / References: UniProt: Q9HAW0
#20: Protein Transcription factor TFIIIB component B'' homolog / Transcription factor IIIB 150 / TFIIIB150 / Transcription factor-like nuclear regulator


Mass: 55103.477 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: BDP1, KIAA1241, KIAA1689, TFNR / Production host: Escherichia coli (E. coli) / References: UniProt: A6H8Y1

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SnRNA-activating protein complex subunit ... , 3 types, 3 molecules UWZ

#21: Protein snRNA-activating protein complex subunit 1 / SNAPc subunit 1 / Proximal sequence element-binding transcription factor subunit gamma / PSE- ...SNAPc subunit 1 / Proximal sequence element-binding transcription factor subunit gamma / PSE-binding factor subunit gamma / PTF subunit gamma / Small nuclear RNA-activating complex polypeptide 1 / snRNA-activating protein complex 43 kDa subunit / SNAPc 43 kDa subunit


Mass: 43074.473 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: SNAPC1, SNAP43 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q16533
#22: Protein snRNA-activating protein complex subunit 4 / SNAPc subunit 4 / Proximal sequence element-binding transcription factor subunit alpha / PSE- ...SNAPc subunit 4 / Proximal sequence element-binding transcription factor subunit alpha / PSE-binding factor subunit alpha / PTF subunit alpha / snRNA-activating protein complex 190 kDa subunit / SNAPc 190 kDa subunit


Mass: 165124.812 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: SNAPC4, SNAP190 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q5SXM2
#25: Protein snRNA-activating protein complex subunit 3 / SNAPc subunit 3 / Proximal sequence element-binding transcription factor subunit beta / PSE-binding ...SNAPc subunit 3 / Proximal sequence element-binding transcription factor subunit beta / PSE-binding factor subunit beta / PTF subunit beta / Small nuclear RNA-activating complex polypeptide 3 / snRNA-activating protein complex 50 kDa subunit / SNAPc 50 kDa subunit


Mass: 46812.605 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: SNAPC3, SNAP50 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q92966

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DNA chain , 2 types, 2 molecules XY

#23: DNA chain U6_2_Template


Mass: 30177.291 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Homo sapiens (human)
#24: DNA chain U6_2_Non template


Mass: 30280.418 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Homo sapiens (human)

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Non-polymers , 3 types, 11 molecules

#26: Chemical
ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 9 / Source method: obtained synthetically / Formula: Zn
#27: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Mg
#28: Chemical ChemComp-SF4 / IRON/SULFUR CLUSTER


Mass: 351.640 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Fe4S4

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Details

Has ligand of interestN
Has protein modificationN

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

Component
IDNameTypeEntity IDParent-IDSource
1RNA Polymerase III Class III Melting Pre-Initiation ComplexCOMPLEX#1-#250MULTIPLE SOURCES
2SNAPcCOMPLEX#21-#22, #251RECOMBINANT
3TFIIIBCOMPLEX#18-#201RECOMBINANT
4DNACOMPLEX#23-#241RECOMBINANT
5Human RNA Polymerase IIICOMPLEX#1-#171NATURAL
Molecular weightExperimental value: NO
Source (natural)
IDEntity assembly-IDOrganismNcbi tax-ID
22Homo sapiens (human)9606
33Homo sapiens (human)9606
44Homo sapiens (human)9606
55Homo sapiens (human)9606
Source (recombinant)
IDEntity assembly-IDOrganismNcbi tax-ID
22Trichoplusia ni (cabbage looper)7111
33Escherichia coli (E. coli)562
44synthetic construct (others)32630
Buffer solutionpH: 8
Buffer component
IDConc.NameBuffer-ID
125 mMHEPES1
2150 mMSodium Chloride1
35 mMMagnisium Chloride1
43 mMDTT1
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen supportGrid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS / Details: FEI Titan Krios G4 (operating at 300 KeV)
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal magnification: 130000 X / Nominal defocus max: 1500 nm / Nominal defocus min: 750 nm / Cs: 2.7 mm
Specimen holderSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image recordingAverage exposure time: 5.3 sec. / Electron dose: 50 e/Å2 / Film or detector model: TFS FALCON 4i (4k x 4k) / Num. of real images: 17291
EM imaging opticsEnergyfilter name: TFS Selectris X / Energyfilter slit width: 10 eV

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Processing

EM software
IDNameVersionCategory
4CTFFIND4.1.4CTF correction
9cryoSPARC4.3.1initial Euler assignment
10RELION4initial Euler assignment
11cryoSPARC4.3.1final Euler assignment
12RELION4final Euler assignment
13cryoSPARC4.3.1classification
14cryoSPARC4.3.13D reconstruction
15PHENIXdev_5246model refinement
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
SymmetryPoint symmetry: C1 (asymmetric)
3D reconstructionResolution: 3.51 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 9476 / Symmetry type: POINT

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