2020-11-18RELPDBePDBePDBe2018-03-222018-04-112018-11-212020-11-18Wellcome Trust101908/Z/13/ZUnited KingdomEuropean Research CouncilERC-CoG-648432 MEMBRANEFUSIONUnited KingdomCryoEM structure of the MDA5-dsRNA filament with 91-degree helical twistYu QQu KModis YYu QQu KModis YCryo-EM Structures of MDA5-dsRNA Filaments at Different Stages of ATP Hydrolysis.Mol. CellUS729991012.e6201830449722doi:10.1016/j.molcel.2018.10.0121097-41642168MOCEFLEMD-4340other EM volumeMDA5-dsRNA filament with 87 degree twistEMD-4338other EM volumeMDA5-dsRNA filament with 74 degree twist and AMPPNP boundEMD-4341associated EM volumeCryoEM structure of the MDA5-dsRNA filament with 91-degree helical twist6g1xFULLOVERLAPMDA5-dsRNA helical filamentMDA5-dsRNA helical filament01232.22MDA511DExD/H-box helicase consisting of Hel1, Hel2, Hel2i, and pincer domains, followed by a C-terminal domainMus musculusEscherichia coli BL21(DE3)pET28aDouble-stranded RNA from bacteriophage Phi6123Pseudomonas phage phi6Pseudomonas savastanoi pv. phaseolicolaInterferon-induced helicase C domain-containing protein 1Mus musculus0.0800576951Escherichia coli BL21(DE3)LEVOLQLRPYQMEVAQPALDGKNIIICLPTGSGKTRVAVYITKDHLDKKKQASESGKVIVLVNKVMLAEQLFRKEFNPYLKKWY
RIIGLSGDTQLKISFPEVVKSYDVIISTAQILENSLLNLESGDDDGVQLSDFSLIIIDECHHTNKEAVYNNIMRRYLKQK
LRNNDLKKQNKPAIPLPQILGLTASPGVGAAKKQSEAEKHILNICANLDAFTIKTVKENLGQLKHQIKEPCKKFVIADDT
RENPFKEKLLEIMASIQTYCQKSPMSDFGTQHYEQWAIQMEKKAAKDGNRKDRVCAEHLRKYNEALQINDTIRMIDAYSH
LETFYTDEKEKKFAVLNDSKKSLKLDETDEFLMNLFFDNKKMLKKLAENPKYENEKLIKLRNTILEQFTRSEESSRGIIF
TKTRQSTYALSQWIMENAKFAEVGVKAHHLIGAGHSSEVKPMTQTEQKEVISKFRTGEINLLIATTVAEEGLDIKECNIV
IRYGLVTNEIAMVQARGRARADESTYVLVTSSGSGVTEREIVNDFREKMMYKAINRVQNMKPEEYAHKILELQVQSILEK
KMKVKRSIAKQYNDNPSLITLLCKNCSMLVCSGENIHVIEKMHHVNMTPEFKGLYIVRENKALQKKFADYQTNGEIICKC
GQAWGTMMVHKGLDLPCLKIRNFVVNFKNNSPKKQYKKWVELPIRFPDLDYSEYCL3.6.4.13RNA (5'-R(P*UP*CP*CP*AP*UP*GP*CP*GP*CP*AP*UP*GP*AP*CP*G)-3')Pseudomonas phage phi60.0047678891UCCAUGCGCAUGACGRNA (5'-R(P*CP*GP*UP*CP*AP*UP*GP*CP*GP*CP*AP*UP*GP*GP*A)-3')Pseudomonas phage phi60.0048079141CGUCAUGCGCAUGGAZINC ION6.5409e-051ZNhelicalhelicalArray0.47.720.0HEPES0.1KClpotassium chloride5.0MgCl2magnesium chloride2.0DTTQuantifoil R1.2/1.3GOLD300CARBONHOLEYGLOW DISCHARGEAIR25 mAETHANE100277FEI VITROBOT MARK IVFilament samples were diluted twofold from 0.8 mg/ml to 0.4 mg/ml immediately prior to plunge freezingFEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN3001.82.775000.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENFEI FALCON III (4k x 4k)COUNTING40964096315630.829.851144.970390.9214C1FOURIER SPACE3.93FSC 0.143 CUT-OFFRELION2.1.039987Gctf1.06367549RELION2.1.0A cylinder was used as the initial model for 3D image reconstructionNOT APPLICABLERELION2.1.0FLEXIBLE FITCross-correlation coefficientREAL165emd_4341_msk_1.mapemd_4341_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
224
224224
0
00224224224239.68001239.68001239.6800190.090.090.0XYZ-0.0216499920.064243590.000383033160.0045111221.071.071.07Half map 1 used in helical reconstruction in RELION 2.1.0emd_4341_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
224
224224
0
00224224224239.68001239.68001239.6800190.090.090.0XYZ-0.0197886250.064399080.00037810240.0045047121.071.071.07Half map 2 used in helical reconstruction in RELION 2.1.0