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- EMDB-42304: Structure of Pol II-DSIF-NELF - post-translocated -

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Basic information

Entry
Database: EMDB / ID: EMD-42304
TitleStructure of Pol II-DSIF-NELF - post-translocated
Map data
Sample
  • Complex: RNA Polymerase in complex with DSIF and NELF
    • Protein or peptide: x 18 types
    • DNA: x 2 types
    • RNA: x 1 types
KeywordsNucleic acids / transcription / RNA polymerase II / NELF / DSIF / pausing
Biological speciesSus scrofa (pig) / Homo sapiens (human) / synthetic construct (others)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.8 Å
AuthorsSu BG / Vos SM
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)DP2-GM146254 United States
CitationJournal: Mol Cell / Year: 2024
Title: Distinct negative elongation factor conformations regulate RNA polymerase II promoter-proximal pausing.
Authors: Bonnie G Su / Seychelle M Vos /
Abstract: Metazoan gene expression regulation involves pausing of RNA polymerase (Pol II) in the promoter-proximal region of genes and is stabilized by DSIF and NELF. Upon depletion of elongation factors, NELF ...Metazoan gene expression regulation involves pausing of RNA polymerase (Pol II) in the promoter-proximal region of genes and is stabilized by DSIF and NELF. Upon depletion of elongation factors, NELF appears to accompany elongating Pol II past pause sites; however, prior work indicates that NELF prevents Pol II elongation. Here, we report cryoelectron microscopy structures of Pol II-DSIF-NELF complexes with NELF in two distinct conformations corresponding to paused and poised states. The paused NELF state supports Pol II stalling, whereas the poised NELF state enables transcription elongation as it does not support a tilted RNA-DNA hybrid. Further, the poised NELF state can accommodate TFIIS binding to Pol II, allowing for Pol II reactivation at paused or backtracking sites. Finally, we observe that the NELF-A tentacle interacts with the RPB2 protrusion and is necessary for pausing. Our results define how NELF can support pausing, reactivation, and elongation by Pol II.
History
DepositionOct 10, 2023-
Header (metadata) releaseMar 20, 2024-
Map releaseMar 20, 2024-
UpdateApr 17, 2024-
Current statusApr 17, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_42304.png

Downloads & links

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Map

FileDownload / File: emd_42304.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

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Size
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AxesZ (Sec.)Y (Row.)X (Col.)
0.82 Å/pix.
x 512 pix.
= 420.864 Å		0.82 Å/pix.
x 512 pix.
= 420.864 Å		0.82 Å/pix.
x 512 pix.
= 420.864 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 0.822 Å
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Histogram (log scale)
Contour LevelBy AUTHOR: 0.35
Minimum - Maximum-0.8710356 - 2.7886388
Average (Standard dev.)0.00023841269 (±0.089455046)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions512512512
Spacing512512512
CellA=B=C: 420.864 Å
α=β=γ: 90.0 °

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Supplemental data

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Mask #1

Fileemd_42304_msk_1.map
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Additional map: #1

Fileemd_42304_additional_1.map
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Half map: #1

Fileemd_42304_half_map_1.map
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Half map: #2

Fileemd_42304_half_map_2.map
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Sample components

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Entire : RNA Polymerase in complex with DSIF and NELF

EntireName: RNA Polymerase in complex with DSIF and NELF
Components
  • Complex: RNA Polymerase in complex with DSIF and NELF
    • Protein or peptide: RPB1
    • Protein or peptide: RPB2
    • Protein or peptide: RPB3
    • Protein or peptide: RPB4
    • Protein or peptide: RPB5
    • Protein or peptide: RPB6
    • Protein or peptide: RPB7
    • Protein or peptide: RPB8
    • Protein or peptide: RPB9
    • Protein or peptide: RPB10
    • Protein or peptide: RPB11
    • Protein or peptide: RPB12
    • DNA: Non-template DNA
    • RNA: RNA
    • DNA: Template DNA
    • Protein or peptide: NELF-A
    • Protein or peptide: SPT4
    • Protein or peptide: SPT5
    • Protein or peptide: NELF-C
    • Protein or peptide: NELF-B
    • Protein or peptide: NELF-E

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Supramolecule #1: RNA Polymerase in complex with DSIF and NELF

SupramoleculeName: RNA Polymerase in complex with DSIF and NELF / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#21

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Macromolecule #1: RPB1

MacromoleculeName: RPB1 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString: MHGGGPPSGD SACPLRTIKR VQFGVLSPDE LKRMSVTEGG IKYPETTEGG RPKLGGLMDP RQGVIERTGR CQTCAGNMTE CPGHFGHIE LAKPVFHVGF LVKTMKVLRC VCFFCSKLLV DSNNPKIKDI LAKSKGQPKK RLTHVYDLCK GKNICEGGEE M DNKFGVEQ ...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 SYSPTSPNYS PTSPNYTPTS P SYSPTSPS YSPTSPNYTP TSPNYSPTSP SYSPTSPSYS PTSPSYSPSS PRYTPQSPTY TPSSPSYSPS SPSYSPTSPK YT PTSPSYS PSSPEYTPTS PKYSPTSPKY SPTSPKYSPT SPTYSPTTPK YSPTSPTYSP TSPVYTPTSP KYSPTSPTYS PTS PKYSPT SPTYSPTSPK GSTYSPTSPG YSPTSPTYSL TSPAISPDDS DEEN

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Macromolecule #2: RPB2

MacromoleculeName: RPB2 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString: MYDADEDMQY DEDDDEITPD LWQEACWIVI SSYFDEKGLV RQQLDSFDEF IQMSVQRIVE DAPPIDLQAE AQHASGEVEE PPRYLLKFE QIYLSKPTHW ERDGAPSPMM PNEARLRNLT YSAPLYVDIT KTVIKEGEEQ LQTQHQKTFI GKIPIMLRST Y CLLNGLTD ...String:
MYDADEDMQY DEDDDEITPD LWQEACWIVI SSYFDEKGLV RQQLDSFDEF IQMSVQRIVE DAPPIDLQAE AQHASGEVEE PPRYLLKFE QIYLSKPTHW ERDGAPSPMM PNEARLRNLT YSAPLYVDIT KTVIKEGEEQ LQTQHQKTFI GKIPIMLRST Y CLLNGLTD RDLCELNECP LDPGGYFIIN GSEKVLIAQE KMATNTVYVF AKKDSKYAYT GECRSCLENS SRPTSTIWVS ML ARGGQGA KKSAIGQRIV ATLPYIKQEV PIIIVFRALG FVSDRDILEH IIYDFEDPEM MEMVKPSLDE AFVIQEQNVA LNF IGSRGA KPGVTKEKRI KYAKEVLQKE MLPHVGVSDF CETKKAYFLG YMVHRLLLAA LGRRELDDRD HYGNKRLDLA GPLL AFLFR GMFKNLLKEV RIYAQKFIDR GKDFNLELAI KTRIISDGLK YSLATGNWGD QKKAHQARAG VSQVLNRLTF ASTLS HLRR LNSPIGRDGK LAKPRQLHNT LWGMVCPAET PEGHAVGLVK NLALMAYISV GSQPSPILEF LEEWSMENLE EISPAA IAD ATKIFVNGCW VGIHKDPEQL MNTLRKLRRQ MDIIVSEVSM IRDIREREIR IYTDAGRICR PLLIVEKQKL LLKKRHI DQ LKEREYNNYS WQDLVASGVV EYIDTLEEET VMLAMTPDDL QEKEVAYCST YTHCEIHPSM ILGVCASIIP FPDHNQSP R NTYQSAMGKQ AMGVYITNFH VRMDTLAHVL YYPQKPLVTT RSMEYLRFRE LPAGINSIVA IASYTGYNQE DSVIMNRSA VDRGFFRSVF YRSYKEQESK KGFDQEEVFE KPTRETCQGM RHAIYDKLDD DGLIAPGVRV SGDDVIIGKT VTLPENEDEL EGTNRRYTK RDCSTFLRTS ETGIVDQVMV TLNQEGYKFC KIRVRSVRIP QIGDKFASRH GQKGTCGIQY RQEDMPFTCE G ITPDIIIN PHAIPSRMTI GHLIECLQGK VSANKGEIGD ATPFNDAVNV QKISNLLSDY GYHLRGNEVL YNGFTGRKIT SQ IFIGPTY YQRLKHMVDD KIHSRARGPI QILNRQPMEG RSRDGGLRFG EMERDCQIAH GAAQFLRERL FEASDPYQVH VCN LCGIMA IANTRTHTYE CRGCRNKTQI SLVRMPYACK LLFQELMSMS IAPRMMSV

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Macromolecule #3: RPB3

MacromoleculeName: RPB3 / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString: MPYANQPTVR ITELTDENVK FIIENTDLAV ANSIRRVFIA EVPIIAIDWV QIDANSSVLH DEFIAHRLGL IPLTSDDIVD KLQYSRDCT CEEFCPECSV EFTLDVRCNE DQTRHVTSRD LISNSPRVIP VTSRNRDNDP SDYVEQDDIL IVKLRKGQEL R LRAYAKKG ...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 QSDV

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Macromolecule #4: RPB4

MacromoleculeName: RPB4 / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MAAGGSDPRA GDVEEDASQL IFPKEFETAE TLLNSEVHML LEHRKQQNES AEDEQELSEV FMKTLNYTAR FSRFKNRETI ASVRSLLLQ KKLHKFELAC LANLCPETAE ESKALIPSLE GRFEDEELQQ ILDDIQTKRS FQY

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Macromolecule #5: RPB5

MacromoleculeName: RPB5 / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString: MDDEEETYRL WKIRKTIMQL CHDRGYLVTQ DELDQTLEEF KAQFGDKPSE GRPRRTDLTV LVAHNDDPTD QMFVFFPEEP KVGIKTIKV YCQRMQEENI TRALIVVQQG MTPSAKQSLV DMAPKYILEQ FLQQELLINI TEHELVPEHV VMTKEEVTEL L ARYKLREN ...String:
MDDEEETYRL WKIRKTIMQL CHDRGYLVTQ DELDQTLEEF KAQFGDKPSE GRPRRTDLTV LVAHNDDPTD QMFVFFPEEP KVGIKTIKV YCQRMQEENI TRALIVVQQG MTPSAKQSLV DMAPKYILEQ FLQQELLINI TEHELVPEHV VMTKEEVTEL L ARYKLREN QLPRIQAGDP VARYFGIKRG QVVKIIRPSE TAGRYITYRL VQ

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Macromolecule #6: RPB6

MacromoleculeName: RPB6 / type: protein_or_peptide / ID: 6 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MSDNEDNFDG DDFDDVEEDE GLDDLENAEE EGQENVEILP SGERPQANQK RITTPYMTKY ERARVLGTRA LQIAMCAPVM VELEGETDP LLIAMKELKA RKIPIIIRRY LPDGSYEDWG VDELIISD

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Macromolecule #7: RPB7

MacromoleculeName: RPB7 / type: protein_or_peptide / ID: 7 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MFYHISLEHE ILLHPRYFGP NLLNTVKQKL FTEVEGTCTG KYGFVIAVTT IDNIGAGVIQ PGRGFVLYPV KYKAIVFRPF KGEVVDAVV TQVNKVGLFT EIGPMSCFIS RHSIPSEMEF DPNSNPPCYK TMDEDIVIQQ DDEIRLKIVG TRVDKNDIFA I GSLMDDYL GLVS

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Macromolecule #8: RPB8

MacromoleculeName: RPB8 / type: protein_or_peptide / ID: 8 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MAGILFEDIF DVKDIDPEGK KFDRVSRLHC ESESFKMDLI LDVNIQIYPV DLGDKFRLVI ASTLYEDGTL DDGEYNPTDD RPSRADQFE YVMYGKVYRI EGDETSTEAA TRLSAYVSYG GLLMRLQGDA NNLHGFEVDS RVYLLMKKLA F

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Macromolecule #9: RPB9

MacromoleculeName: RPB9 / type: protein_or_peptide / ID: 9 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MEPDGTYEPG FVGIRFCQEC NNMLYPKEDK ENRILLYACR NCDYQQEADN SCIYVNKITH EVDELTQIIA DVSQDPTLPR TEDHPCQKC GHKEAVFFQS HSARAEDAMR LYYVCTAPHC GHRWTE

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Macromolecule #10: RPB10

MacromoleculeName: RPB10 / type: protein_or_peptide / ID: 10 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MIIPVRCFTC GKIVGNKWEA YLGLLQAEYT EGDALDALGL KRYCCRRMLL AHVDLIEKLL NYAPLEK

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Macromolecule #11: RPB11

MacromoleculeName: RPB11 / type: protein_or_peptide / ID: 11 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MNAPPAFESF LLFEGEKKIT INKDTKVPNA CLFTINKEDH TLGNIIKSQL LKDPQVLFAG YKVPHPLEHK IIIRVQTTPD YSPQEAFTN AITDLISELS LLEERFRVAI KDKQEGIE

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Macromolecule #12: RPB12

MacromoleculeName: RPB12 / type: protein_or_peptide / ID: 12 / Enantiomer: LEVO
Source (natural)Organism: Sus scrofa (pig)
SequenceString:
MDTQKDVQPP KQQPMIYICG ECHTENEIKS RDPIRCRECG YRIMYKKRTK RLVVFDAR

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Macromolecule #16: NELF-A

MacromoleculeName: NELF-A / type: protein_or_peptide / ID: 16 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MASMRESDTG LWLHNKLGAT DELWAPPSIA SLLTAAVIDN IRLCFHGLSS AVKLKLLLGT LHLPRRTVDE MKGALMEIIQ LASLDSDPWV LMVADILKSF PDTGSLNLEL EEQNPNVQDI LGELREKVGE CEASAMLPLE CQYLNKNALT TLAGPLTPPV KHFQLKRKPK ...String:
MASMRESDTG LWLHNKLGAT DELWAPPSIA SLLTAAVIDN IRLCFHGLSS AVKLKLLLGT LHLPRRTVDE MKGALMEIIQ LASLDSDPWV LMVADILKSF PDTGSLNLEL EEQNPNVQDI LGELREKVGE CEASAMLPLE CQYLNKNALT TLAGPLTPPV KHFQLKRKPK SATLRAELLQ KSTETAQQxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xxxxxxxxxx xGVKLLDISE LDMVGAGREA KRRRKTLDAE VVEKPAKEET VVENATPDYA AGLVSTQKLG SLNNEPALPS TSYLPSTPSV VPASSYIPSS ETPPAPSSRE ASRPPEEPSA PSPTLPAQFK QRAPMYNSGL SPATPTPAAP TSPLTPTTPP AVAPTTQTPP VAMVAPQTQA PAQQQPKKNL SLTREQMFAA QEMFKTANKV TRPEKALILG FMAGSRENPC QEQGDVIQIK LSEHTEDLPK ADGQGSTTML VDTVFEMNYA TGQWTRFKKY KPMTNVS

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Macromolecule #17: SPT4

MacromoleculeName: SPT4 / type: protein_or_peptide / ID: 17 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString:
GPGSMALETV PKDLRHLRAC LLCSLVKTID QFEYDGCDNC DAYLQMKGNR EMVYDCTSSS FDGIIAMMSP EDSWVSKWQR VSNFKPGVY AVSVTGRLPQ GIVRELKSRG VAYKSRDTAI KT

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Macromolecule #18: SPT5

MacromoleculeName: SPT5 / type: protein_or_peptide / ID: 18 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MSDSEDSNFS EEEDSERSSD GEEAEVDEER RSAAGSEKEE EPEDEEEEEE EEEYDEEEEE EDDDRPPKKP RHGGFILDEA DVDDEYEDE DQWEDGAEDI LEKEEIEASN IDNVVLDEDR SGARRLQNLW RDQREEELGE YYMKKYAKSS VGETVYGGSD E LSDDITQQ ...String:
MSDSEDSNFS EEEDSERSSD GEEAEVDEER RSAAGSEKEE EPEDEEEEEE EEEYDEEEEE EDDDRPPKKP RHGGFILDEA DVDDEYEDE DQWEDGAEDI LEKEEIEASN IDNVVLDEDR SGARRLQNLW RDQREEELGE YYMKKYAKSS VGETVYGGSD E LSDDITQQ QLLPGVKDPN LWTVKCKIGE ERATAISLMR KFIAYQFTDT PLQIKSVVAP EHVKGYIYVE AYKQTHVKQA IE GVGNLRL GYWNQQMVPI KEMTDVLKVV KEVANLKPKS WVRLKRGIYK DDIAQVDYVE PSQNTISLKM IPRIDYDRIK ARM SLKDWF AKRKKFKRPP QRLFDAEKIR SLGGDVASDG DFLIFEGNRY SRKGFLFKSF AMSAVITEGV KPTLSELEKF EDQP EGIDL EVVTESTGKE REHNFQPGDN VEVCEGELIN LQGKILSVDG NKITIMPKHE DLKDMLEFPA QELRKYFKMG DHVKV IAGR FEGDTGLIVR VEENFVILFS DLTMHELKVL PRDLQLCSET ASGVDVGGQH EWGELVQLDP QTVGVIVRLE RETFQV LNM YGKVVTVRHQ AVTRKKDNRF AVALDSEQNN IHVKDIVKVI DGPHSGREGE IRHLFRSFAF LHCKKLVENG GMFVCKT RH LVLAGGSKPR DVTNFTVGGF APMSPRISSP MHPSAGGQRG GFGSPGGGSG GMSRGRGRRD NELIGQTVRI SQGPYKGY I GVVKDATEST ARVELHSTCQ TISVDRQRLT TVGSRRPGGM TSTYGRTPMY GSQTPMYGSG SRTPMYGSQT PLQDGSRTP HYGSQTPLHD GSRTPAQSGA WDPNNPNTPS RAEEEYEYAF DDEPTPSPQA YGGTPNPQTP GYPDPSSPQV NPQYNPQTPG TPAMYNTDQ FSPYAAPSPQ GSYQPSPSPQ SYHQVAPSPA GYQNTHSPAS YHPTPSPMAY QASPSPSPVG YSPMTPGAPS P GGYNPHTP GSGIEQNSSD WVTTDIQVKV RDTYLDTQVV GQTGVIRSVT GGMCSVYLKD SEKVVSISSE HLEPITPTKN NK VKVILGE DREATGVLLS IDGEDGIVRM DLDEQLKILN LRFLGKLLEA

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Macromolecule #19: NELF-C

MacromoleculeName: NELF-C / type: protein_or_peptide / ID: 19 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: SNAMDEDYYG SAAEWGDEAD GGQQEDDSGE GEDDAEVQQE CLHKFSTRDY IMEPSIFNTL KRYFQAGGSP ENVIQLLSEN YTAVAQTVN LLAEWLIQTG VEPVQVQETV ENHLKSLLIK HFDPRKADSI FTEEGETPAW LEQMIAHTTW RDLFYKLAEA H PDCLMLNF ...String:
SNAMDEDYYG SAAEWGDEAD GGQQEDDSGE GEDDAEVQQE CLHKFSTRDY IMEPSIFNTL KRYFQAGGSP ENVIQLLSEN YTAVAQTVN LLAEWLIQTG VEPVQVQETV ENHLKSLLIK HFDPRKADSI FTEEGETPAW LEQMIAHTTW RDLFYKLAEA H PDCLMLNF TVKLISDAGY QGEITSVSTA CQQLEVFSRV LRTSLATILD GGEENLEKNL PEFAKMVCHG EHTYLFAQAM MS VLAQEEQ GGSAVRRIAQ EVQRFAQEKG HDASQITLAL GTAASYPRAC QALGAMLSKG ALNPADITVL FKMFTSMDPP PVE LIRVPA FLDLFMQSLF KPGARINQDH KHKYIHILAY AASVVETWKK NKRVSINKDE LKSTSKAVET VHNLCCNENK GASE LVAEL STLYQCIRFP VVAMGVLKWV DWTVSEPRYF QLQTDHTPVH LALLDEISTC HQLLHPQVLQ LLVKLFETEH SALDV MEQL ELKKTLLDRM VHLLSRGYVL PVVSYIRKCL EKLDTDISLI RYFVTEVLDV IAPPYTSDFV QLFLPILEND SIAGTI KTE GEHDPVTEFI AHCKSNFIMV N

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Macromolecule #20: NELF-B

MacromoleculeName: NELF-B / type: protein_or_peptide / ID: 20 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MFAGLQDLGV ANGEDLKETL TNCTEPLKAI EQFQTENGVL LPSLQSALPF LDLHGTPRLE FHQSVFDELR DKLLERVSAI ASEGKAEER YKKLEDLLEK SFSLVKMPSL QPVVMCVMKH LPKVPEKKLK LVMADKELYR ACAVEVKRQI WQDNQALFGD E VSPLLKQY ...String:
MFAGLQDLGV ANGEDLKETL TNCTEPLKAI EQFQTENGVL LPSLQSALPF LDLHGTPRLE FHQSVFDELR DKLLERVSAI ASEGKAEER YKKLEDLLEK SFSLVKMPSL QPVVMCVMKH LPKVPEKKLK LVMADKELYR ACAVEVKRQI WQDNQALFGD E VSPLLKQY ILEKESALFS TELSVLHNFF SPSPKTRRQG EVVQRLTRMV GKNVKLYDMV LQFLRTLFLR TRNVHYCTLR AE LLMSLHD LDVGEICTVD PCHKFTWCLD ACIRERFVDS KRARELQGFL DGVKKGQEQV LGDLSMILCD PFAINTLALS TVR HLQELV GQETLPRDSP DLLLLLRLLA LGQGAWDMID SQVFKEPKME VELITRFLPM LMSFLVDDYT FNVDQKLPAE EKAP VSYPN TLPESFTKFL QEQRMACEVG LYYVLHITKQ RNKNALLRLL PGLVETFGDL AFGDIFLHLL TGNLALLADE FALED FCSS LFDGFFLTAS PRKENVHRHA LRLLIHLHPR VAPSKLEALQ KALEPTGQSG EAVKELYSQL GEKLEQLDHR KPSPAQ AAE TPALELPLPS VPAPAPL

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Macromolecule #21: NELF-E

MacromoleculeName: NELF-E / type: protein_or_peptide / ID: 21 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MLVIPPGMSE EEEALQKKFM KLKKKKKALM ALKKQSSSST TSQGGVKRSL SEQPVMDTAT ATEQAKQLVK SGAISAIKAE TKNSGFKRS RTLEGKLKDP EKGPVPTFQP FQRSISADDD LQESSRRPQR KSLYESFVSS SDRLRELGPD GEEAEGPGAG D GPPRSFDW ...String:
MLVIPPGMSE EEEALQKKFM KLKKKKKALM ALKKQSSSST TSQGGVKRSL SEQPVMDTAT ATEQAKQLVK SGAISAIKAE TKNSGFKRS RTLEGKLKDP EKGPVPTFQP FQRSISADDD LQESSRRPQR KSLYESFVSS SDRLRELGPD GEEAEGPGAG D GPPRSFDW GYEERSGAHS SASPPRSRSR DRSHERNRDR DRDRERDRDR DRDRDRERDR DRDRDRDRDR ERDRDRERDR DR DREGPFR RSDSFPERRA PRKGNTLYVY GEDMTPTLLR GAFSPFGNII DLSMDPPRNC AFVTYEKMES ADQAVAELNG TQV ESVQLK VNIARKQPML DAATGKSVWG SLAVQNSPKG CHRDKRTQIV YSDDVYKENL VDGF

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Macromolecule #13: Non-template DNA

MacromoleculeName: Non-template DNA / type: dna / ID: 13 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
SequenceString:
(DG)(DG)(DT)(DC)(DA)(DG)(DT)(DA)(DC)(DG) (DT)(DC)(DC)(DG)(DG)(DC)(DA)(DT)(DA)(DG) (DT)(DT)(DG)(DC)(DG)(DC)(DC)(DC)(DG) (DT)(DA)(DA)(DA)(DT)(DT)(DC)(DA)(DG)(DA) (DT) (DC)(DT)(DT)(DC)(DC)(DA)(DG)(DT) (DG)(DG)

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Macromolecule #15: Template DNA

MacromoleculeName: Template DNA / type: dna / ID: 15 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
SequenceString:
(DC)(DC)(DA)(DC)(DT)(DG)(DG)(DA)(DA)(DG) (DA)(DT)(DC)(DT)(DG)(DA)(DA)(DT)(DT)(DT) (DA)(DC)(DG)(DG)(DG)(DC)(DG)(DC)(DA) (DA)(DC)(DT)(DA)(DT)(DG)(DC)(DC)(DG)(DG) (DA) (DC)(DG)(DT)(DA)(DC)(DT)(DG)(DA) (DC)(DC)

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Macromolecule #14: RNA

MacromoleculeName: RNA / type: rna / ID: 14
Source (natural)Organism: synthetic construct (others)
SequenceString:
UUUUUUGGCA UAGUUGCGCC CG

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 51.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination 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

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Image processing

Startup modelType of model: EMDB MAP
EMDB ID:

0038

Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 50535
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD

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