EMDB-19726: RNA polymerase II early elongation complex bound to TFIIE and TFI...

Basic information

Entry

Database: EMDB / ID: EMD-19726

• Title: RNA polymerase II early elongation complex bound to TFIIE and TFIIF - state a (composite structure)
• Map data: Composite map for model building of EC14a (sharpened)

Sample

• Complex: RNA polymerase II early elongation complex bound to TFIIE and TFIIF - state a
  • Complex: RNA polymerase II
    • Protein or peptide: x 12 types
  • Complex: DNA and RNA
    • DNA: x 2 types
    • RNA: x 1 types
  • Cell: Transcription factor IIE and IIF
    • Protein or peptide: x 4 types
• Ligand: x 2 types

• Keywords: RNA polymerase II / promoter escape / de novo transcription / TRANSCRIPTION

Function / homology

Function:

transcription factor TFIIE complex / phosphatase activator activity / TFIIF-class transcription factor complex binding / transcription factor TFIIF complex / Formation of RNA Pol II elongation complex / Formation of the Early Elongation Complex / Transcriptional regulation by small RNAs / RNA Polymerase II Pre-transcription Events / TP53 Regulates Transcription of DNA Repair Genes / FGFR2 alternative splicing / RNA polymerase II transcribes snRNA genes / mRNA Capping / mRNA Splicing - Minor Pathway / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Elongation / RNA Polymerase II Transcription Initiation And Promoter Clearance / RNA Pol II CTD phosphorylation and interaction with CE / Estrogen-dependent gene expression / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA Splicing - Major Pathway / nuclear lumen / RNA polymerase II general transcription initiation factor binding / Abortive elongation of HIV-1 transcript in the absence of Tat / FGFR2 alternative splicing / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / RNA polymerase II general transcription initiation factor activity / transcription factor TFIID complex / 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 / 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 / mRNA Splicing - Minor Pathway / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / Processing of Capped Intron-Containing Pre-mRNA / RNA polymerase II transcribes snRNA genes / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / transcription by RNA polymerase III / transcription by RNA polymerase I / RNA polymerase I complex / transcription elongation by RNA polymerase I / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase III complex / transcription-coupled nucleotide-excision repair / RNA polymerase II, core complex / tRNA transcription by RNA polymerase III / RNA Polymerase II Transcription Elongation / : / Formation of RNA Pol II elongation complex / translation initiation factor binding / RNA Polymerase II Pre-transcription Events / negative regulation of protein binding / DNA-directed RNA polymerase activity / DNA-directed RNA polymerase complex / mRNA Splicing - Major Pathway / positive regulation of transcription elongation by RNA polymerase II / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / DNA-templated transcription initiation / transcription elongation by RNA polymerase II / promoter-specific chromatin binding / response to virus / ribonucleoside binding / : / : / : / fibrillar center / : / : / : / DNA-directed RNA polymerase / cell junction / microtubule cytoskeleton / protein phosphatase binding / Estrogen-dependent gene expression / transcription by RNA polymerase II / nucleic acid binding / forked DNA-dependent helicase activity / single-stranded 3'-5' DNA helicase activity / four-way junction helicase activity / double-stranded DNA helicase activity / chromosome, telomeric region

Domain/homology:

Transcription factor TFIIE alpha subunit, C-terminal / C-terminal general transcription factor TFIIE alpha / Transcription factor TFIIE beta subunit, DNA-binding domain / Transcription initiation factor TFIIE, beta subunit / TFA2, Winged helix domain 2 / TFIIE beta subunit core domain / TFA2 Winged helix domain 2 / TFIIE beta central core DNA-binding domain profile. / Transcription initiation factor IIE subunit alpha, N-terminal / Transcription factor TFE/TFIIEalpha HTH domain / TFIIEalpha/SarR/Rpc3 HTH domain / Transcription factor E / TFIIE alpha subunit / TFE/IIEalpha-type HTH domain profile. / Transcription initiation factor IIE / Transcription initiation factor IIF, beta subunit / TFIIF beta subunit, HTH domain / TFIIF, beta subunit, N-terminal / TFIIF, beta subunit HTH domain / TFIIF, beta subunit N-terminus / Transcription initiation factor IIF, alpha subunit / Transcription initiation factor IIF, alpha subunit (TFIIF-alpha) / Transcription Factor IIF, Rap30/Rap74, interaction / Zinc finger, TFIIB-type / TFIIB zinc-binding / DNA-directed RNA polymerase II subunit Rpb4-like / RNA polymerase Rpb1 C-terminal repeat / RNA polymerase II, heptapeptide repeat, eukaryotic / Eukaryotic RNA polymerase II heptapeptide repeat. / RNA polymerase Rpb1, domain 6 / RNA polymerase Rpb1, domain 6 / RNA polymerase Rpb4/RPC9, core / DNA-directed RNA-polymerase II subunit / RNA polymerase Rpb1, domain 7 / RNA polymerase Rpb1, domain 7 superfamily / RNA polymerase Rpb1, domain 7 / Pol II subunit B9, C-terminal zinc ribbon / RNA polymerase RBP11 / Rpb4/RPC9 superfamily / RNA polymerase subunit Rpb4/RPC9 / RNA polymerase Rpb4 / 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 / RNA polymerases L / 13 to 16 Kd subunits signature. / Zinc finger, TFIIS-type / Transcription factor S-II (TFIIS) / Zinc finger TFIIS-type profile. / C2C2 Zinc finger / RNA polymerase subunit RPABC4/transcription elongation factor Spt4 / DNA-directed RNA polymerase, RBP11-like dimerisation domain / RNA polymerase Rpb3/Rpb11 dimerisation domain / RNA polymerase, subunit H/Rpb5 C-terminal / DNA-directed RNA polymerase subunit Rpo5/Rpb5 / RPB5-like RNA polymerase subunit superfamily / RNA polymerase Rpb5, C-terminal domain / Archaeal Rpo6/eukaryotic RPB6 RNA polymerase subunit / DNA-directed RNA polymerase, 14-18kDa subunit, conserved site / RNA polymerases K / 14 to 18 Kd subunits signature. / Ribosomal protein S1-like RNA-binding domain / S1 RNA binding domain / S1 domain / DNA-directed RNA polymerase, subunit beta-prime

Component:

RNA polymerase II subunit D / DNA-directed RNA polymerase subunit / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / DNA-directed RNA polymerase subunit / RNA polymerase II subunit K / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerase II subunit RPB11-a / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / DNA-directed RNA polymerase II subunit RPB3 / DNA-directed RNA polymerase subunit beta / DNA-directed RNA polymerase II subunit E / General transcription factor IIF subunit 2 / General transcription factor IIE subunit 1 / Transcription initiation factor IIE subunit beta / General transcription factor IIF subunit 1 / DNA-directed RNA polymerase II subunit RPB9

• Biological species: Sus scrofa (pig) / unidentified adenovirus / Homo sapiens (human)
• Method: single particle reconstruction / cryo EM / Resolution: 3.4 Å
• Authors: Zhan Y / Grabbe F / Oberbeckmann E / Dienemann C / Cramer P

Funding support

Germany, 1 items

Organization	Grant number	Country
Max Planck Society		Germany

• Citation: Journal: Mol Cell / Year: 2024; Title: Three-step mechanism of promoter escape by RNA polymerase II.; Authors: Yumeng Zhan / Frauke Grabbe / Elisa Oberbeckmann / Christian Dienemann / Patrick Cramer / ; Abstract: The transition from transcription initiation to elongation is highly regulated in human cells but remains incompletely understood at the structural level. In particular, it is unclear how interactions between RNA polymerase II (RNA Pol II) and initiation factors are broken to enable promoter escape. Here, we reconstitute RNA Pol II promoter escape in vitro and determine high-resolution structures of initially transcribing complexes containing 8-, 10-, and 12-nt ordered RNAs and two elongation complexes containing 14-nt RNAs. We suggest that promoter escape occurs in three major steps. First, the growing RNA displaces the B-reader element of the initiation factor TFIIB without evicting TFIIB. Second, the rewinding of the transcription bubble coincides with the eviction of TFIIA, TFIIB, and TBP. Third, the binding of DSIF and NELF facilitates TFIIE and TFIIH dissociation, establishing the paused elongation complex. This three-step model for promoter escape fills a gap in our understanding of the initiation-elongation transition of RNA Pol II transcription.

History

Deposition	Feb 22, 2024	-
Header (metadata) release	Apr 17, 2024	-
Map release	Apr 17, 2024	-
Update	Nov 6, 2024	-
Current status	Nov 6, 2024	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_19726.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Composite map for model building of EC14a (sharpened)
• Voxel size: X=Y=Z: 1.05 Å

Density

Contour Level	By AUTHOR: 3.37
Minimum - Maximum	-23.055520999999999 - 47.040207000000002
Average (Standard dev.)	-0.000000000001989 (±1.0)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order Z Y X Origin 0 0 0 Dimensions 400 400 400 Spacing 400 400 400
Cell	A=B=C: 419.99997 Å α=β=γ: 90.0 °

Supplemental data

Mask #1

• File: emd_19726_msk_1.map

Additional map: Half map corresponding to the map focused refined on EC14a.

• File: emd_19726_additional_1.map
• Annotation: Half map corresponding to the map focused refined on EC14a.

Additional map: Half map corresponding to the map focused refined on EC14a.

• File: emd_19726_additional_2.map
• Annotation: Half map corresponding to the map focused refined on EC14a.

Additional map: Focused refined map of EC14a (locally filtered).

• File: emd_19726_additional_3.map
• Annotation: Focused refined map of EC14a (locally filtered).

Additional map: Mask for focused refinement on Pol II

• File: emd_19726_additional_4.map
• Annotation: Mask for focused refinement on Pol II

Additional map: Half map corresponding to the map focused refined on Pol II

• File: emd_19726_additional_5.map
• Annotation: Half map corresponding to the map focused refined on Pol II

Additional map: Half map corresponding to the map focused refined on Pol II.

• File: emd_19726_additional_6.map
• Annotation: Half map corresponding to the map focused refined on Pol II.

Additional map: Focused refined map of Pol II (locally filtered).

• File: emd_19726_additional_7.map
• Annotation: Focused refined map of Pol II (locally filtered).

Sample components

Entire : RNA polymerase II early elongation complex bound to TFIIE and TFI...

• Entire: Name: RNA polymerase II early elongation complex bound to TFIIE and TFIIF - state a

Components

• Complex: RNA polymerase II early elongation complex bound to TFIIE and TFIIF - state a
  • Complex: RNA polymerase II
    • Protein or peptide: DNA-directed RNA polymerase subunit
    • Protein or peptide: DNA-directed RNA polymerase subunit beta
    • Protein or peptide: DNA-directed RNA polymerase II subunit RPB3
    • Protein or peptide: RNA polymerase II subunit D
    • Protein or peptide: DNA-directed RNA polymerase II subunit E
    • Protein or peptide: DNA-directed RNA polymerases I, II, and III subunit RPABC2
    • Protein or peptide: DNA-directed RNA polymerase II subunit RPB7
    • Protein or peptide: DNA-directed RNA polymerases I, II, and III subunit RPABC3
    • Protein or peptide: DNA-directed RNA polymerase II subunit RPB9
    • Protein or peptide: DNA-directed RNA polymerases I, II, and III subunit RPABC5
    • Protein or peptide: DNA-directed RNA polymerase II subunit RPB11-a
    • Protein or peptide: RNA polymerase II subunit K
  • Complex: DNA and RNA
    • DNA: Non-template DNA
    • RNA: RNA
    • DNA: Template DNA
  • Cell: Transcription factor IIE and IIF
    • Protein or peptide: General transcription factor IIF subunit 1
    • Protein or peptide: General transcription factor IIF subunit 2
    • Protein or peptide: General transcription factor IIE subunit 1
    • Protein or peptide: Transcription initiation factor IIE subunit beta
• Ligand: ZINC ION
• Ligand: MAGNESIUM ION

Supramolecule #1: RNA polymerase II early elongation complex bound to TFIIE and TFI...

• Supramolecule: Name: RNA polymerase II early elongation complex bound to TFIIE and TFIIF - state a; type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#19

Supramolecule #2: RNA polymerase II

• Supramolecule: Name: RNA polymerase II / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#12
• Source (natural): Organism: Sus scrofa (pig)

Supramolecule #3: DNA and RNA

• Supramolecule: Name: DNA and RNA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #13-#14, #17
• Source (natural): Organism: unidentified adenovirus

Supramolecule #4: Transcription factor IIE and IIF

• Supramolecule: Name: Transcription factor IIE and IIF / type: cell / ID: 4 / Parent: 1 / Macromolecule list: #15-#16, #18-#19
• Source (natural): Organism: Homo sapiens (human)

Macromolecule #1: DNA-directed RNA polymerase subunit

• Macromolecule: Name: DNA-directed RNA polymerase subunit / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 218.889547 KDa

Sequence

String:

MHGGGPPSGD SACPLRTIKR VQFGVLSPDE LKRMSVTEGG IKYPETTEGG RPKLGGLMDP RQGVIERTGR CQTCAGNMTE CPGHFGHIE LAKPVFHVGF LVKTMKVLRC VCFFCSKLLV DSNNPKIKDI LAKSKGQPKK RLTHVYDLCK GKNICEGGEE M DNKFGVEQ PEGDEDLTKE KGHGGCGRYQ PRIRRSGLEL YAEWKHVNED SQEKKILLSP ERVHEIFKRI SDEECFVLGM EP RYARPEW MIVTVLPVPP LSVRPAVVMQ GSARNQDDLT HKLADIVKIN NQLRRNEQNG AAAHVIAEDV KLLQFHVATM VDN ELPGLP RAMQKSGRPL KSLKQRLKGK EGRVRGNLMG KRVDFSARTV ITPDPNLSID QVGVPRSIAA NMTFAEIVTP FNID RLQEL VRRGNSQYPG AKYIIRDNGD RIDLRFHPKP SDLHLQTGYK VERHMCDGDI VIFNRQPTLH KMSMMGHRVR ILPWS TFRL NLSVTTPYNA DFDGDEMNLH LPQSLETRAE IQELAMVPRM IVTPQSNRPV MGIVQDTLTA VRKFTKRDVF LERGEV MNL LMFLSTWDGK VPQPAILKPR PLWTGKQIFS LIIPGHINCI RTHSTHPDDE DSGPYKHISP GDTKVVVENG ELIMGIL CK KSLGTSAGSL VHISYLEMGH DITRLFYSNI QTVINNWLLI EGHTIGIGDS IADSKTYQDI QNTIKKAKQD VIEVIEKA H NNELEPTPGN TLRQTFENQV NRILNDARDK TGSSAQKSLS EYNNFKSMVV SGAKGSKINI SQVIAVVGQQ NVEGKRIPF GFKHRTLPHF IKDDYGPESR GFVENSYLAG LTPTEFFFHA MGGREGLIDT AVKTAETGYI QRRLIKSMES VMVKYDATVR NSINQVVQL RYGEDGLAGE SVEFQNLATL KPSNKAFEKK FRFDYTNERA LRRTLQEDLV KDVLSNAHIQ NELEREFERM R EDREVLRV IFPTGDSKVV LPCNLLRMIW NAQKIFHINP RLPSDLHPIK VVEGVKELSK KLVIVNGDDP LSRQAQENAT LL FNIHLRS TLCSRRMAEE FRLSGEAFDW LLGEIESKFN QAIAHPGEMV GALAAQSLGE PATQMTLNTF HYAGVSAKNV TLG VPRLKE LINISKKPKT PSLTVFLLGQ SARDAERAKD ILCRLEHTTL RKVTANTAIY YDPNPQSTVV AEDQEWVNVY YEMP DFDVA RISPWLLRVE LDRKHMTDRK LTMEQIAEKI NAGFGDDLNC IFNDDNAEKL VLRIRIMNSD ENKMQEEEEV VDKMD DDVF LRCIESNMLT DMTLQGIEQI SKVYMHLPQT DNKKKIIITE DGEFKALQEW ILETDGVSLM RVLSEKDVDP VRTTSN DIV EIFTVLGIEA VRKALERELY HVISFDGSYV NYRHLALLCD TMTCRGHLMA ITRHGVNRQD TGPLMKCSFE ETVDVLM EA AAHGESDPMK GVSENIMLGQ LAPAGTGCFD LLLDAEKCKY GMEIPTNIPG LGAAGPTGMF FGSAPSPMGG ISPAMTPW N QGATPAYGAW SPSVGSGMTP GAAGFSPSAA SDASGFSPGY SPAWSPTPGS PGSPGPSSPY IPSPGGAMSP SYSPTSPAY EPRSPGGYTP QSPSYSPTSP SYSPTSPSYS PTSPNYSPTS PSYSPTSPSY SPTSPSYSPT SPSYSPTSPS YSPTSPSYSP TSPSYSPTS PSYSPTSPSY SPTSPSYSPT SPSYSPTSPS YSPTSPSYSP TSPSYSPTSP SYSPTSPSYS PTSPSYSPTS P NYSPTSPN YTPTSPSYSP TSPSYSPTSP NYTPTSPNYS PTSPSYSPTS PSYSPTSPSY SPSSPRYTPQ SPTYTPSSPS YS PSSPSYS PTSPKYTPTS PSYSPSSPEY TPTSPKYSPT SPKYSPTSPK YSPTSPTYSP TTPKYSPTSP TYSPTSPVYT PTS PKYSPT SPTYSPTSPK YSPTSPTYSP TSPKGSTYSP TSPGYSPTSP TYSLTSPAIS PDDSDEEN

UniProtKB: DNA-directed RNA polymerase subunit

Macromolecule #2: DNA-directed RNA polymerase subunit beta

• Macromolecule: Name: DNA-directed RNA polymerase subunit beta / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 147.938594 KDa

Sequence

String:

MKPLRPENPP KAELKGLLFV QQHCKTRLHS VAVTRPRGAA TEESTPKLRM CSTNLSQALA YFRAGANLTA ASLWAGFRGS RRIFWERRK RKSLCLALRP GGACWRWLLA VSCVSLRGLG APGSCANMYD ADEDMQYDED DDEITPDLWQ EACWIVISSY F DEKGLVRQ QLDSFDEFIQ MSVQRIVEDA PPIDLQAEAQ HASGEVEEPP RYLLKFEQIY LSKPTHWERD GAPSPMMPNE AR LRNLTYS APLYVDITKT VIKEGEEQLQ TQHQKTFIGK IPIMLRSTYC LLNGLTDRDL CELNECPLDP GGYFIINGSE KVL IAQEKM ATNTVYVFAK KDSKYAYTGE CRSCLENSSR PTSTIWVSML ARGGQGAKKS AIGQRIVATL PYIKQEVPII IVFR ALGFV SDRDILEHII YDFEDPEMME MVKPSLDEAF VIQEQNVALN FIGSRGAKPG VTKEKRIKYA KEVLQKEMLP HVGVS DFCE TKKAYFLGYM VHRLLLAALG RRELDDRDHY GNKRLDLAGP LLAFLFRGMF KNLLKEVRIY AQKFIDRGKD FNLELA IKT RIISDGLKYS LATGNWGDQK KAHQARAGVS QVLNRLTFAS TLSHLRRLNS PIGRDGKLAK PRQLHNTLWG MVCPAET PE GHAVGLVKNL ALMAYISVGS QPSPILEFLE EWSMENLEEI SPAAIADATK IFVNGCWVGI HKDPEQLMNT LRKLRRQM D IIVSEVSMIR DIREREIRIY TDAGRICRPL LIVEKQKLLL KKRHIDQLKE REYNNYSWQD LVASGVVEYI DTLEEETVM LAMTPDDLQE KEVAYCSTYT HCEIHPSMIL GVCASIIPFP DHNQSPRNTY QSAMGKQAMG VYITNFHVRM DTLAHVLYYP QKPLVTTRS MEYLRFRELP AGINSIVAIA SYTGYNQEDS VIMNRSAVDR GFFRSVFYRS YKEQESKKGF DQEEVFEKPT R ETCQGMRH AIYDKLDDDG LIAPGVRVSG DDVIIGKTVT LPENEDELEG TNRRYTKRDC STFLRTSETG IVDQVMVTLN QE GYKFCKI RVRSVRIPQI GDKFASRHGQ KGTCGIQYRQ EDMPFTCEGI TPDIIINPHA IPSRMTIGHL IECLQGKVSA NKG EIGDAT PFNDAVNVQK ISNLLSDYGY HLRGNEVLYN GFTGRKITSQ IFIGPTYYQR LKHMVDDKIH SRARGPIQIL NRQP MEGRS RDGGLRFGEM ERDCQIAHGA AQFLRERLFE ASDPYQVHVC NLCGIMAIAN TRTHTYECRG CRNKTQISLV RMPYA CKLL FQELMSMSIA PRMMSV

UniProtKB: DNA-directed RNA polymerase subunit beta

Macromolecule #3: DNA-directed RNA polymerase II subunit RPB3

• Macromolecule: Name: DNA-directed RNA polymerase II subunit RPB3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 31.439074 KDa

Sequence

String:

MPYANQPTVR ITELTDENVK FIIENTDLAV ANSIRRVFIA EVPIIAIDWV QIDANSSVLH DEFIAHRLGL IPLTSDDIVD KLQYSRDCT CEEFCPECSV EFTLDVRCNE DQTRHVTSRD LISNSPRVIP VTSRNRDNDP SDYVEQDDIL IVKLRKGQEL R LRAYAKKG FGKEHAKWNP TAGVAFEYDP DNALRHTVYP KPEEWPKSEY SELDEDESQA PYDPNGKPER FYYNVESCGS LR PETIVLS ALSGLKKKLS DLQTQLSHEI QSDVLTIN

UniProtKB: DNA-directed RNA polymerase II subunit RPB3

Macromolecule #4: RNA polymerase II subunit D

• Macromolecule: Name: RNA polymerase II subunit D / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 20.962621 KDa

Sequence

String:

MWGPAQPYSD SALSPKPRPF RAVFRGSALP FPAVRVEVRG RSMAAGGSDP RAGDVEEDAS QLIFPKEFET AETLLNSEVH MLLEHRKQQ NESAEDEQEL SEVFMKTLNY TARFSRFKNR ETIASVRSLL LQKKLHKFEL ACLANLCPET AEESKALIPS L EGRFEDEE LQQILDDIQT KRSFQY

UniProtKB: RNA polymerase II subunit D

Macromolecule #5: DNA-directed RNA polymerase II subunit E

• Macromolecule: Name: DNA-directed RNA polymerase II subunit E / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 24.644318 KDa

Sequence

String:

MDDEEETYRL WKIRKTIMQL CHDRGYLVTQ DELDQTLEEF KAQFGDKPSE GRPRRTDLTV LVAHNDDPTD QMFVFFPEEP KVGIKTIKV YCQRMQEENI TRALIVVQQG MTPSAKQSLV DMAPKYILEQ FLQQELLINI TEHELVPEHV VMTKEEVTEL L ARYKLREN QLPRIQAGDP VARYFGIKRG QVVKIIRPSE TAGRYITYRL VQ

UniProtKB: DNA-directed RNA polymerase II subunit E

Macromolecule #6: DNA-directed RNA polymerases I, II, and III subunit RPABC2

• Macromolecule: Name: DNA-directed RNA polymerases I, II, and III subunit RPABC2; type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 14.477001 KDa

Sequence

String:

MSDNEDNFDG DDFDDVEEDE GLDDLENAEE EGQENVEILP SGERPQANQK RITTPYMTKY ERARVLGTRA LQIAMCAPVM VELEGETDP LLIAMKELKA RKIPIIIRRY LPDGSYEDWG VDELIISD

UniProtKB: DNA-directed RNA polymerases I, II, and III subunit RPABC2

Macromolecule #7: DNA-directed RNA polymerase II subunit RPB7

• Macromolecule: Name: DNA-directed RNA polymerase II subunit RPB7 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 19.314283 KDa

Sequence

String:

MFYHISLEHE ILLHPRYFGP NLLNTVKQKL FTEVEGTCTG KYGFVIAVTT IDNIGAGVIQ PGRGFVLYPV KYKAIVFRPF KGEVVDAVV TQVNKVGLFT EIGPMSCFIS RHSIPSEMEF DPNSNPPCYK TMDEDIVIQQ DDEIRLKIVG TRVDKNDIFA I GSLMDDYL GLVS

UniProtKB: DNA-directed RNA polymerase subunit

Macromolecule #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3

• Macromolecule: Name: DNA-directed RNA polymerases I, II, and III subunit RPABC3; type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 17.162273 KDa

Sequence

String:

MAGILFEDIF DVKDIDPEGK KFDRVSRLHC ESESFKMDLI LDVNIQIYPV DLGDKFRLVI ASTLYEDGTL DDGEYNPTDD RPSRADQFE YVMYGKVYRI EGDETSTEAA TRLSAYVSYG GLLMRLQGDA NNLHGFEVDS RVYLLMKKLA F

UniProtKB: DNA-directed RNA polymerases I, II, and III subunit RPABC3

Macromolecule #9: DNA-directed RNA polymerase II subunit RPB9

• Macromolecule: Name: DNA-directed RNA polymerase II subunit RPB9 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 14.541221 KDa

Sequence

String:

MEPDGTYEPG FVGIRFCQEC NNMLYPKEDK ENRILLYACR NCDYQQEADN SCIYVNKITH EVDELTQIIA DVSQDPTLPR TEDHPCQKC GHKEAVFFQS HSARAEDAMR LYYVCTAPHC GHRWTE

UniProtKB: DNA-directed RNA polymerase II subunit RPB9

Macromolecule #10: DNA-directed RNA polymerases I, II, and III subunit RPABC5

• Macromolecule: Name: DNA-directed RNA polymerases I, II, and III subunit RPABC5; type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 7.655123 KDa

Sequence

String:

MIIPVRCFTC GKIVGNKWEA YLGLLQAEYT EGDALDALGL KRYCCRRMLL AHVDLIEKLL NYAPLEK

UniProtKB: DNA-directed RNA polymerases I, II, and III subunit RPABC5

Macromolecule #11: DNA-directed RNA polymerase II subunit RPB11-a

• Macromolecule: Name: DNA-directed RNA polymerase II subunit RPB11-a / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 13.310284 KDa

Sequence

String:

MNAPPAFESF LLFEGEKKIT INKDTKVPNA CLFTINKEDH TLGNIIKSQL LKDPQVLFAG YKVPHPLEHK IIIRVQTTPD YSPQEAFTN AITDLISELS LLEERFRVAI KDKQEGIE

UniProtKB: DNA-directed RNA polymerase II subunit RPB11-a

Macromolecule #12: RNA polymerase II subunit K

• Macromolecule: Name: RNA polymerase II subunit K / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Sus scrofa (pig)
• Molecular weight: Theoretical: 7.018244 KDa

Sequence

String:

MDTQKDVQPP KQQPMIYICG ECHTENEIKS RDPIRCRECG YRIMYKKRTK RLVVFDAR

UniProtKB: RNA polymerase II subunit K

Macromolecule #15: General transcription factor IIF subunit 1

• Macromolecule: Name: General transcription factor IIF subunit 1 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 58.343578 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

MAALGPSSQN VTEYVVRVPK NTTKKYNIMA FNAADKVNFA TWNQARLERD LSNKKIYQEE EMPESGAGSE FNRKLREEAR RKKYGIVLK EFRPEDQPWL LRVNGKSGRK FKGIKKGGVT ENTSYYIFTQ CPDGAFEAFP VHNWYNFTPL ARHRTLTAEE A EEEWERRN KVLNHFSIMQ QRRLKDQDQD EDEEEKEKRG RRKASELRIH DLEDDLEMSS DASDASGEEG GRVPKAKKKA PL AKGGRKK KKKKGSDDEA FEDSDDGDFE GQEVDYMSDG SSSSQEEPES KAKAPQQEEG PKGVDEQSDS SEESEEEKPP EED KEEEEE KKAPTPQEKK RRKDSSEESD SSEESDIDSE ASSALFMAKK KTPPKRERKP SGGSSRGNSR PGTPSAEGGS TSST LRAAA SKLEQGKRVS EMPAAKRLRL DTGPQSLSGK STPQPPSGKT TPNSGDVQVT EDAVRRYLTR KPMTTKDLLK KFQTK KTGL SSEQTVNVLA QILKRLNPER KMINDKMHFS LKE

UniProtKB: General transcription factor IIF subunit 1

Macromolecule #16: General transcription factor IIF subunit 2

• Macromolecule: Name: General transcription factor IIF subunit 2 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 28.427309 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

MAERGELDLT GAKQNTGVWL VKVPKYLSQQ WAKASGRGEV GKLRIAKTQG RTEVSFTLNE DLANIHDIGG KPASVSAPRE HPFVLQSVG GQTLTVFTES SSDKLSLEGI VVQRAECRPA ASENYMRLKR LQIEESSKPV RLSQQLDKVV TTNYKPVANH Q YNIEYERK KKEDGKRARA DKQHVLDMLF SAFEKHQYYN LKDLVDITKQ PVVYLKEILK EIGVQNVKGI HKNTWELKPE YR HYQGEEK SD

UniProtKB: General transcription factor IIF subunit 2

Macromolecule #18: General transcription factor IIE subunit 1

• Macromolecule: Name: General transcription factor IIE subunit 1 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 49.516094 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

MADPDVLTEV PAALKRLAKY VIRGFYGIEH ALALDILIRN SCVKEEDMLE LLKFDRKQLR SVLNNLKGDK FIKCRMRVET AADGKTTRH NYYFINYRTL VNVVKYKLDH MRRRIETDER DSTNRASFKC PVCSSTFTDL EANQLFDPMT GTFRCTFCHT E VEEDESAM PKKDARTLLA RFNEQIEPIY ALLRETEDVN LAYEILEPEP TEIPALKQSK DHAATTAGAA SLAGGHHREA WA TKGPSYE DLYTQNVVIN MDDQEDLHRA SLEGKSAKER PIWLRESTVQ GAYGSEDMKE GGIDMDAFQE REEGHAGPDD NEE VMRALL IHEKKTSSAM AGSVGAAAPV TAANGSDSES ETSESDDDSP PRPAAVAVHK REEDEEEDDE FEEVADDPIV MVAG RPFSY SEVSQRPELV AQMTPEEKEA YIAMGQRMFE DLFE

UniProtKB: General transcription factor IIE subunit 1

Macromolecule #19: Transcription initiation factor IIE subunit beta

• Macromolecule: Name: Transcription initiation factor IIE subunit beta / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 33.106824 KDa
• Recombinant expression: Organism: Escherichia coli (E. coli)

Sequence

String:

MDPSLLRERE LFKKRALSTP VVEKRSASSE SSSSSSKKKK TKVEHGGSSG SKQNSDHSNG SFNLKALSGS SGYKFGVLAK IVNYMKTRH QRGDTHPLTL DEILDETQHL DIGLKQKQWL MTEALVNNPK IEVIDGKYAF KPKYNVRDKK ALLRLLDQHD Q RGLGGILL EDIEEALPNS QKAVKALGDQ ILFVNRPDKK KILFFNDKSC QFSVDEEFQK LWRSVTVDSM DEEKIEEYLK RQ GISSMQE SGPKKVAPIQ RRKKPASQKK RRFKTHNEHL AGVLKDYSDI TSSK

UniProtKB: Transcription initiation factor IIE subunit beta

Macromolecule #13: Non-template DNA

• Macromolecule: Name: Non-template DNA / type: dna / ID: 13 / Number of copies: 1 / Classification: DNA
• Source (natural): Organism: unidentified adenovirus
• Molecular weight: Theoretical: 43.236555 KDa

Sequence

String:

(DG)(DT)(DG)(DT)(DT)(DC)(DC)(DT)(DG)(DA) (DA)(DG)(DG)(DG)(DG)(DG)(DG)(DC)(DT)(DA) (DT)(DA)(DA)(DA)(DA)(DG)(DG)(DG)(DG) (DG)(DT)(DG)(DG)(DG)(DG)(DG)(DC)(DG)(DC) (DG) (DT)(DT)(DC)(DG)(DT)(DC)(DC)(DT) (DC)(DA)(DC)(DA)(DC)(DC)(DC)(DA)(DG)(DC) (DC)(DG) (DC)(DA)(DA)(DT)(DT)(DT)(DT) (DG)(DA)(DC)(DA)(DG)(DC)(DG)(DA)(DG)(DG) (DG)(DC)(DC) (DA)(DG)(DC)(DA)(DG)(DA) (DA)(DG)(DG)(DG)(DG)(DA)(DG)(DA)(DG)(DA) (DA)(DC)(DT)(DT) (DT)(DT)(DT)(DC)(DT) (DC)(DA)(DA)(DA)(DA)(DG)(DC)(DG)(DG)(DG) (DC)(DA)(DT)(DG)(DA) (DC)(DT)(DT)(DC) (DT)(DG)(DC)(DG)(DC)(DT)(DA)(DA)(DG)(DA) (DT)(DT)(DG)(DT)(DC)

Macromolecule #17: Template DNA

• Macromolecule: Name: Template DNA / type: dna / ID: 17 / Number of copies: 1 / Classification: DNA
• Source (natural): Organism: unidentified adenovirus
• Molecular weight: Theoretical: 42.569098 KDa

Sequence

String:

(DG)(DA)(DC)(DA)(DA)(DT)(DC)(DT)(DT)(DA) (DG)(DC)(DG)(DC)(DA)(DG)(DA)(DA)(DG)(DT) (DC)(DA)(DT)(DG)(DC)(DC)(DC)(DG)(DC) (DT)(DT)(DT)(DT)(DG)(DA)(DG)(DA)(DA)(DA) (DA) (DA)(DG)(DT)(DT)(DC)(DT)(DC)(DT) (DC)(DC)(DC)(DC)(DT)(DT)(DC)(DT)(DG)(DC) (DT)(DG) (DG)(DC)(DC)(DC)(DT)(DC)(DG) (DC)(DT)(DG)(DT)(DC)(DA)(DA)(DA)(DA)(DT) (DT)(DG)(DC) (DG)(DG)(DC)(DT)(DG)(DG) (DG)(DT)(DG)(DT)(DG)(DA)(DG)(DG)(DA)(DC) (DG)(DA)(DA)(DC) (DG)(DC)(DG)(DC)(DC) (DC)(DC)(DC)(DA)(DC)(DC)(DC)(DC)(DC)(DT) (DT)(DT)(DT)(DA)(DT) (DA)(DG)(DC)(DC) (DC)(DC)(DC)(DC)(DT)(DT)(DC)(DA)(DG)(DG) (DA)(DA)(DC)(DA)(DC)

Macromolecule #14: RNA

• Macromolecule: Name: RNA / type: rna / ID: 14 / Number of copies: 1
• Source (natural): Organism: unidentified adenovirus
• Molecular weight: Theoretical: 4.427756 KDa

Sequence

String:

ACACCCAGCC GCAA

Macromolecule #20: ZINC ION

• Macromolecule: Name: ZINC ION / type: ligand / ID: 20 / Number of copies: 9 / Formula: ZN
• Molecular weight: Theoretical: 65.409 Da

Macromolecule #21: MAGNESIUM ION

• Macromolecule: Name: MAGNESIUM ION / type: ligand / ID: 21 / Number of copies: 1 / Formula: MG
• Molecular weight: Theoretical: 24.305 Da

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

• Buffer: pH: 7.5
• Vitrification: Cryogen name: ETHANE

Electron microscopy

• Microscope: FEI TITAN KRIOS
• Image recording: Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 40.0 e/Ų
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.7 µm / Nominal defocus min: 0.7000000000000001 µm
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

Startup model

Type of model: EMDB MAP
EMDB ID:

12613

• Final reconstruction: Resolution.type: BY AUTHOR / Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF; Details: Number of particles is different for every focused refined map, as detailed in the manuscript.; Number images used: 50000
• Initial angle assignment: Type: MAXIMUM LIKELIHOOD
• Final angle assignment: Type: MAXIMUM LIKELIHOOD