2024-11-13 REL PDBe PDBe PDBe 2020-02-05 2020-11-25 2020-11-25 2024-11-13 Wellcome Trust 200835/Z/16/Z United Kingdom Medical Research Council (MRC, United Kingdom) MR/R009945/1 United Kingdom Medical Research Council (MRC, United Kingdom) MR/K000241/1 United Kingdom Fan H Carrique L Influenza Polymerase, ANP32, replication, RNA-dependent RNA polymerase, VIRAL PROTEIN Carrique L Fan H Walker AP Keown JR Sharps J Staller E Barclay WS Fodor E Grimes JM Nature UK 587 638 643 2020 33208942 doi:10.1038/s41586-020-2927-z 1476-4687 0006 NATUAS 6xzq FULLOVERLAP Influenza C virus polymerase in complex with human ANP32A Influenza C virus polymerase in complex with human ANP32A 0 1 2 3 4 5 Influenza C virus (C/Johannesburg/1/66) Influenza C virus polymerases 1 2 3 4 Influenza C virus (C/Johannesburg/1/66) Human Acidic leucine-rich nuclear phosphoprotein 32 family member A (ANP32A) 1 5 Homo sapiens Influenza viral RNA (vRNA) promoter 47mer 1 1

Synthetic RNA

synthetic construct RNA (5'-R(*AP*GP*UP*AP*GP*AP*AP*AP*CP*AP*AP*GP*GP*GP*CP*CP*CP*UP*G)-3') synthetic construct 0.014901736999999998 1 AGUAGAAACAAGGGUAUUUUUCUUUACUAGUCUACCCUGCUUUUGCU Polymerase acidic protein Influenza C virus (strain C/Johannesburg/1/1966) 0.082025531 2 Spodoptera frugiperda LEVO MSKTFAEIAEAFLEPEAVRIAKEAVEEYGDHERKIIQIGIHFQVCCMFCDEYLSTNGSDRFVLIEGRKRGTAVSLQNELC KSYDLEPLPFLCDIFDREEKQFVEIGITRKADDSYFQSKFGKLGNSCKIFVFSYDGRLDKNCEGPMEEQKLRIFSFLATA ADFLRKENMFNEIFLPDNEETIIEMKKGKTFLELRDESVPLPFQTYEQMKDYCEKFKGNPRELASKVSQMQSNIKLPIKH YEQNKFRQIRLPKGPMAPYTHKFLMEEAWMFTKISDPERSRAGEILIDFFKKGNLSAIRPKDKPLQGKYPIHYKNLWNQI KAAIADRTMVINENDHSEFLGGIGRASKKIPEISLTQDVITTEGLKQSENKLPEPRSFPRWFNAEWMWAIKDSDLTGWVP MAEYPPADNELEDYAEHLNKTMEGVLQGTNCAREMGKCILTVGALMTECRLFPGKIKVVPIYARSKERKSMQEGLPVPSE MDCLFGICVKSKSHLNKDDGMYTIITFEFSIREPNLEKHQKYTVFEAGHTTVRMKKGESVIGREVPLYLYCRTTALSKIK NDWLSKARRCFITTMDTVETICLRESAKAEENLVEKTLNEKQMWIGKKNGELIAQPLREALRVQLVQQFYFCIYNDSQLE GFCNEQKKILMALEGDKKNKSSFGFNPEGLLEKIEECLINNPMCLFMAQRLNELVIEASKRGAKFFKTD Q9IMP5 3.1.-.- RNA-directed RNA polymerase catalytic subunit Influenza C virus (strain C/Johannesburg/1/1966) 0.086145945 2 Spodoptera frugiperda LEVO MEINPYLMFLNNDVTSLISTTYPYTGPPPMSHGSSTKYTLETIKRTYDYSRTSVEKTSKVFNIPRRKFCNCLEDKDELVK PTGNVDISSLLGLAEMMEKRMGEGFFKHCVMEAETEILKMHFSRLTEGRQTYDWTSERNMPAATALQLTVDAIKETEGPF KGTTMLEYCNKMIEMLDWKEIKFKKVKTVVRREKDKRSGKEIKTKVPVMGIDSIKHDEFLIRALTINTMAKDGERGKLQR RAIATPGMIVRPFSKIVETVAQKICEKLKESGLPVGGNEKKAKLKTTVTSLNARMNSDQFAVNITGDNSKWNECQQPEAY LALLAYITKDSSDLMKDLCSVAPVLFCNKFVKLGQGIRLSNKRKTKEVIIKAEKMGKYKNLMREEYKNLFEPLEKYIQKD VCFLPGGMLMGMFNMLSTVLGVSTLCYMDEELKAKGCFWTGLQSSDDFVLFAVASNWSNIHWTIRRFNAVCKLIGINMSL EKSYGSLPELFEFTSMFFDGEFVSNLAMELPAFTTAGVNEGVDFTAAMSIIKTNMINNSLSPSTALMALRICLQEFRATY RVHPWDSRVKGGRMKIINEFIKTIENKDGLLIADGGKLMNNISTLHIPEEVLKFEKMDEQYRNRVFNPKNPFTNFDKTID IFRAHGPIRVEENEAVVSTHSFRTRANRTLLNTDMRAMMAEEKRYQMVCDMFKSVFESADINPPIGAMSIGEAIEEKLLE RAKMKRDIGAIEDSEYEEIKDIIRDAKKARLESR Q9IMP4 2.7.7.48 Polymerase basic protein 2 Influenza C virus (strain C/Johannesburg/1/1966) 0.104272359 2 Spodoptera frugiperda LEVO MSLLLTIAKEYKRLCQDAKAAQMMTVGTVSNYTTFKKWTTSRKEKNPSLRMRWAMSSKFPIIANKRMLEEAQIPKEHNNV ALWEDTEDVSKRDHVLASASCINYWNFCGPCVNNSEVIKEVYKSRFGRLERRKEIMWKELRFTLVDRQRRRVDTQPVEQR LRTGEIKDLQMWTLFEDEAPLASKFILDNYGLVKEMRSKFANKPLNKEVVAHMLEKQFNPESRFLPVFGAIRPERMELIH ALGGETWIQEANTAGISNVDQRKNDIRAVCRKVCLAANASIMNAKSKLVEYIKSTSMRIGETERKLEELILETDDVSPEV TLCKSALGGQLGKTLSFGPMLLKKISGSGVKVKDTVYIQGVRAVQFEYWSEQEEFYGEYKSATALFSRKERSLEWITIGG GINEDRKRLLAMCMIFCRDGDYFKDAPATITMADLSTKLGREIPYQYVMMNWIQKSEDNLEALLYSRGIVETNPGKMGSS MGIDGSKRAIKSLRAVTIQSGKIDMPESKEKIHLELSDNLEAFDSSGRIVATILDLPSDKKVTFQDVSFQHPDLAVLRDE KTAITKGYEALIKRLGTGDNDIPSLIAKKDYLSLYNLPEVKLMAPLIRPNRKGVYSRVARKLVSTQVTTGHYSLHELIKV LPFTYFAPKQGMFEGRLFFSNDSFVEPGVNNNVFSWSKADSSKIYCHGIAIRVPLVVGDEHMDTSLALLEGFSVCENDPR APMVTRQDLIDVGFGQKVRLFVGQGSVRTFKRTASQRAASSDVNKNVKKIKMSNENLYFQGELKTAALAQHDEAVDNKFN KEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRN AFIQSLKADPSQSANLLAEAKKLNGAQAPKVDANSAGKST Q9IMP3 Acidic leucine-rich nuclear phosphoprotein 32 family member A Homo sapiens 0.030328221 1 Spodoptera frugiperda LEVO HHHHHHLEVLFEGPMEMGRRIHLELRNRTPSDVKELVLDNSRSNEGKLEGLTDEFEELEFLSTINVGLTSIANLPKLNKL KKLELSDNRVSGGLEVLAEKCPNLTHLNLSGNKIKDLSTIEPLKKLENLKSLDLFNCEVTNLNDYRENVFKLLPQLTYLD GYDRDDKEAPDSDAEGYVEGLDDEEEDEDEEEYDEDAQVVEDEEDEDEEEEGEEEDVSGEEEEDEEGYNDGEVDDEEDEE ELGEEERGQKRKREPEDEGEDDD P39687 singleParticle particle 0.28 7.5 25.0 C8H18N2O4S HEPES 150.0 NaCL sodium chloride 0.005 C58H114O26 Tween 20 ETHANE 100 294 FEI VITROBOT MARK IV FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 50.0 2.7 2.0 3.5 130000.0 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GIF Bioquantum 20 GATAN K3 (6k x 4k) SUPER-RESOLUTION 1-30 1 9287 2.0 32.1

Single specimen tilt of 20 degrees for around three quarter of the images

1 2312045 1 C1 3.6 FSC 0.143 CUT-OFF cryoSPARC 2.12 85000 ANGULAR RECONSTITUTION cryoSPARC 2.12 MAXIMUM LIKELIHOOD cryoSPARC 2.12 3 80000.0 cryoSPARC 2.12 RIGID BODY FIT

The structure has been determined by single particle cryo-EM. PDB ids 4XOS, 5D98, and 6RR7 were used as initial models after rigid body fitting in chimera.

REAL emd_10665_additional_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 300 300 300 0 0 0 300 300 300 325.5 325.5 325.5 90.0 90.0 90.0 X Y Z -1.7505113 3.3661387 0.0041789147 0.09501721 1.085 1.085 1.085 AUTHOR ::::EMDATABANK.org::::EMD-10665:::: RELION 3.1 locally filtered map emd_10665_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 300 300 300 0 0 0 300 300 300 325.5 325.5 325.5 90.0 90.0 90.0 X Y Z -1.7844323 1.6384497 0.0023449317 0.18537624 1.085 1.085 1.085 AUTHOR ::::EMDATABANK.org::::EMD-10665:::: emd_10665_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 300 300 300 0 0 0 300 300 300 325.5 325.5 325.5 90.0 90.0 90.0 X Y Z -1.6621063 1.5637466 0.0023208624 0.18546839 1.085 1.085 1.085 AUTHOR ::::EMDATABANK.org::::EMD-10665::::