2019-12-11RELPDBePDBePDBe2018-01-262018-02-282018-03-072019-12-11European Research CouncilERC starting grant 679734FranceCryoEM structure of E.coli RNA polymerase paused elongation complex without RNA hairpin bound to NusAGuo XWeixlbaumer AGuo XMyasnikov AGChen JCrucifix CPapai GTakacs MSchultz PWeixlbaumer AStructural Basis for NusA Stabilized Transcriptional Pausing.Mol. CellUS69816827.e4201829499136doi:10.1016/j.molcel.2018.02.0081097-41642168MOCEFLEMD-4274associated EM volumeCryoEM structure of E.coli RNA polymerase paused elongation complex without RNA hairpin bound to NusA6flpFULLOVERLAPCryo-EM structure of E. coli RNA polymerase paused elongation complex without RNA hairpin bound to NusACryo-EM structure of E. coli RNA polymerase paused elongation complex without RNA hairpin bound to NusA01234567RNA polymerase and NusA11234Escherichia coli K-12Escherichia coli BL21(DE3)Nucleic acids1567synthetic constructsynthetic constructDNA-directed RNA polymerase subunit alphaEscherichia coli (strain K12)K120.036558682Escherichia coli BL21(DE3)LEVOMQGSVTEFLKPRLVDIEQVSSTHAKVTLEPLERGFGHTLGNALRRILLSSMPGCAVTEVEIDGVLHEYSTKEGVQEDILE
ILLNLKGLAVRVQGKDEVILTLNKSGIGPVTAADITHDGDVEIVKPQHVICHLTDENASISMRIKVQRGRGYVPASTRIH
SEEDERPIGRLLVDACYSPVERIAYNVEAARVEQRTDLDKLVIEMETNGTIDPEEAIRRAATILAEQLEAFVDLRDVRQP
EVKEEKPEFDPILLRPVDDLELTVRSANCLKAEAIHYIGDLVQRTEVELLKTPNLGKKSLTEIKDVLASRGLSLGMRLEN
WPPASIADE2.7.7.6DNA-directed RNA polymerase subunit betaEscherichia coli (strain K12)K120.1508208751Escherichia coli BL21(DE3)LEVOMVYSYTEKKRIRKDFGKRPQVLDVPYLLSIQLDSFQKFIEQDPEGQYGLEAAFRSVFPIQSYSGNSELQYVSYRLGEPVF
DVQECQIRGVTYSAPLRVKLRLVIYEREAPEGTVKDIKEQEVYMGEIPLMTDNGTFVINGTERVIVSQLHRSPGVFFDSD
KGKTHSSGKVLYNARIIPYRGSWLDFEFDPKDNLFVRIDRRRKLPATIILRALNYTTEQILDLFFEKVIFEIRDNKLQME
LVPERLRGETASFDIEANGKVYVEKGRRITARHIRQLEKDDVKLIEVPVEYIAGKVVAKDYIDESTGELICAANMELSLD
LLAKLSQSGHKRIETLFTNDLDHGPYISETLRVDPTNDRLSALVEIYRMMRPGEPPTREAAESLFENLFFSEDRYDLSAV
GRMKFNRSLLREEIEGSGILSKDDIIDVMKKLIDIRNGKGEVDDIDHLGNRRIRSVGEMAENQFRVGLVRVERAVKERLS
LGDLDTLMPQDMINAKPISAAVKEFFGSSQLSQFMDQNNPLSEITHKRRISALGPGGLTRERAGFEVRDVHPTHYGRVCP
IETPEGPNIGLINSLSVYAQTNEYGFLETPYRKVTDGVVTDEIHYLSAIEEGNYVIAQANSNLDEEGHFVEDLVTCRSKG
ESSLFSRDQVDYMDVSTQQVVSVGASLIPFLEHDDANRALMGANMQRQAVPTLRADKPLVGTGMERAVAVDSGVTAVAKR
GGVVQYVDASRIVIKVNEDEMYPGEAGIDIYNLTKYTRSNQNTCINQMPCVSLGEPVERGDVLADGPSTDLGELALGQNM
RVAFMPWNGYNFEDSILVSERVVQEDRFTTIHIQELACVSRDTKLGPEEITADIPNVGEAALSKLDESGIVYIGAEVTGG
DILVGKVTPKGETQLTPEEKLLRAIFGEKASDVKDSSLRVPNGVSGTVIDVQVFTRDGVEKDKRALEIEEMQLKQAKKDL
SEELQILEAGLFSRIRAVLVAGGVEAEKLDKLPRDRWLELGLTDEEKQNQLEQLAEQYDELKHEFEKKLEAKRRKITQGD
DLAPGVLKIVKVYLAVKRRIQPGDKMAGRHGNKGVISKINPIEDMPYDENGTPVDIVLNPLGVPSRMNIGQILETHLGMA
AKGIGDKINAMLKQQQEVAKLREFIQRAYDLGADVRQKVDLSTFSDEEVMRLAENLRKGMPIATPVFDGAKEAEIKELLK
LGDLPTSGQIRLYDGRTGEQFERPVTVGYMYMLKLNHLVDDKMHARSTGSYSLVTQQPLGGKAQFGGQRFGEMEVWALEA
YGAAYTLQEMLTVKSDDVNGRTKMYKNIVDGNHQMEPGMPESFNVLLKEIRSLGINIELEDE2.7.7.6DNA-directed RNA polymerase subunit beta'Escherichia coli (strain K12)K120.1553667811Escherichia coli BL21(DE3)LEVOMKDLLKFLKAQTKTEEFDAIKIALASPDMIRSWSFGEVKKPETINYRTFKPERDGLFCARIFGPVKDYECLCGKYKRLKH
RGVICEKCGVEVTQTKVRRERMGHIELASPTAHIWFLKSLPSRIGLLLDMPLRDIERVLYFESYVVIEGGMTNLERQQIL
TEEQYLDALEEFGDEFDAKMGAEAIQALLKSMDLEQECEQLREELNETNSETKRKKLTKRIKLLEAFVQSGNKPEWMILT
VLPVLPPDLRPLVPLDGGRFATSDLNDLYRRVINRNNRLKRLLDLAAPDIIVRNEKRMLQEAVDALLDNGRRGRAITGSN
KRPLKSLADMIKGKQGRFRQNLLGKRVDYSGRSVITVGPYLRLHQCGLPKKMALELFKPFIYGKLELRGLATTIKAAKKM
VEREEAVVWDILDEVIREHPVLLNRAPTLHRLGIQAFEPVLIEGKAIQLHPLVCAAYNADFDGDQMAVHVPLTLEAQLEA
RALMMSTNNILSPANGEPIIVPSQDVVLGLYYMTRDCVNAKGEGMVLTGPKEAERLYRSGLASLHARVKVRITEYEKDAN
GELVAKTSLKDTTVGRAILWMIVPKGLPYSIVNQALGKKAISKMLNTCYRILGLKPTVIFADQIMYTGFAYAARSGASVG
IDDMVIPEKKHEIISEAEAEVAEIQEQFQSGLVTAGERYNKVIDIWAAANDRVSKAMMDNLQTETVINRDGQEEKQVSFN
SIYMMADSGARGSAAQIRQLAGMRGLMAKPDGSIIETPITANFREGLNVLQYFISTHGARKGLADTALKTANSGYLTRRL
VDVAQDLVVTEDDCGTHEGIMMTPVIEGGDVKEPLRDRVLGRVTAEDVLKPGTADILVPRNTLLHEQWCDLLEENSVDAV
KVRSVVSCDTDFGVCAHCYGRDLARGHIINKGEAIGVIAAQSIGEPGTQLTMRTFHIGGAASRAAAESSIQVKNKGSIKL
SNVKSVVNSSGKLVITSRNTELKLIDEFGRTKESYKVPYGAVLAKGDGEQVAGGETVANWDPHTMPVITEVSGFVRFTDM
IDGQTITRQTDELTGLSSLVVLDSAERTAGGKDLRPALKIVDAQGNDVLIPGTDMPAQYFLPGKAIVQLEDGVQISSGDT
LARIPQESGGTKDITGGLPRVADLFEARRPKEPAILAEISGIVSFGKETKGKRRLVITPVDGSDPYEEMIPKWRQLNVFE
GERVERGDVISDGPEAPHDILRLRGVHAVTRYIVNEVQDVYRLQGVKINDKHIEVIVRQMLRKATIVNAGSSDFLEGEQV
EYSRVKIANRELEANGKVGATYSRDLLGITKASLATESFISAASFQETTRVLTEAAVAGKRDELRGLKENVIVGRLIPAG
TGYAYHQDRMRRRAAGEAPAAPQVTAEDASASLAELLNAGLGGSDNE2.7.7.6DNA-directed RNA polymerase subunit omegaEscherichia coli (strain K12)K120.0102495471Escherichia coli BL21(DE3)LEVOMARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDKTTVIALREIEEGLINNQILDVRERQEQQEQEAAEL
QAVTAIAEGRR2.7.7.6DNA (30-MER)synthetic construct0.0096762351(DG)(DG)(DG)(DA)(DC)(DG)(DT)(DA)(DC)(DT)(DG)(DA)(DC)(DC)(DG)(DC)(DG)(DG)(DA)(DA)
(DG)(DA)(DG)(DA)(DT)(DT)(DC)(DA)(DG)(DA)(DG)DNARNA (5'-R(P*GP*AP*UP*GP*UP*GP*UP*GP*CP*U)-3')synthetic construct0.0031949171GAUGUGUGCUDNA (39-MER)synthetic construct0.0118155861(DC)(DT)(DC)(DT)(DG)(DA)(DA)(DT)(DC)(DT)(DC)(DT)(DT)(DC)(DC)(DA)(DG)(DC)(DA)(DC)
(DA)(DC)(DA)(DT)(DC)(DG)(DG)(DT)(DC)(DA)(DG)(DT)(DA)(DC)(DG)(DT)(DC)(DC)(DC)DNAMAGNESIUM ION2.4305e-051MGZINC ION6.5409e-052ZNsingleParticleparticle810.0C8H18N2O4SHEPES150.0CH3CO2KPotassium acetate2.0C4H10O2S2Dithiothreitol10.0ZnCl2Zinc chloride5.0Mg(CH3COO)2Magnesium acetate8.0C32H58N2O8SCHAPSOC-flat-1.2/1.3 4CCOPPER400ETHANE95283.15FEI VITROBOT MARK IVFEI TITAN KRIOSSPOT SCANBRIGHT FIELDFIELD EMISSION GUN3000.0010.83.2105000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENGIF Quantum ER020GATAN K2 SUMMIT (4k x 4k)SUPER-RESOLUTION3-4049578.053.0165966CTFFIND44YG2C14.1FSC 0.143 CUT-OFFcryoSPARC65966PROJECTION MATCHINGRELION2.1PROJECTION MATCHINGcryoSPARCRELION2.1RIGID BODY FITREALemd_4274_msk_1.mapemd_4274_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
280
280280
0
00280280280308.0308.0308.090.090.090.0XYZ-1.02690121.0774630.00438442970.130980861.11.11.1emd_4274_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)