2025-09-03 REL PDBe INITIAL_RELEASE REPOSITORY DATABASE_REFERENCES citation citation_author em_admin _citation.country _citation.journal_abbrev _citation.journal_id_ASTM _citation.journal_id_CSD _citation.journal_id_ISSN _citation.journal_volume _citation.page_first _citation.page_last _citation.pdbx_database_id_PubMed _citation.title _citation_author.identifier_ORCID _citation_author.name _em_admin.last_update DATABASE_REFERENCES citation em_admin _citation.journal_volume _citation.page_first _citation.page_last _em_admin.last_update PDBe PDBe 2024-10-04 2025-07-30 2025-07-30 2025-09-03 Research Foundation - Flanders (FWO) 1162823N Belgium Goovaerts Q Shen J Ajjugal Y De Wijngaert B Patel SS Das K POLRMT, TFB2M, TFAM, transcription initiation, polymerase, h-mtRNAP, mtRNAP, human mitochondrial RNA polymerase, y-mtRNAP, MTF1, RPO41, DNA transcription, mitochondrial DNA, mitochondria, TRANSCRIPTION Shen J Goovaerts Q Ajjugal Y De Wijngaert B Das K Patel SS Mol.Cell US 85 3137 3150.e7 2025 40712586 doi:10.1016/j.molcel.2025.07.002 1097-2765 2168 MOCEFL EMD-51727 other EM volume EMD-51729 other EM volume EMD-53853 other EM volume EMD-53854 other EM volume 9gzn FULLOVERLAP EMDB EMD-51728 associated EM volume

Cryo-EM structure of IC3 without TFAM. A second state of the human mitochondrial RNA polymerase transcription initiation complex with 2-mer RNA and GTP poised for catalysis (IC3-TFAM)

EMDB EMD-51727 other EM volume EMDB EMD-51729 other EM volume EMDB EMD-53853 other EM volume EMDB EMD-53854 other EM volume PDB 9GZM unspecified PDB 9GZO unspecified PDB 9R95 unspecified PDB 9R96 unspecified Cryo-EM structure of the human mitochondrial RNA polymerase transcription initiation complex (POLRMT/TFB2M/DNA/RNA) without TFAM; and with a 2-mer RNA (pppGpA) and GTP poised for catalysis (pre-IC3-TFAM) Cryo-EM structure of the human mitochondrial RNA polymerase transcription initiation complex (POLRMT/TFB2M/DNA/RNA) without TFAM; and with a 2-mer RNA (pppGpA) and GTP poised for catalysis (pre-IC3-TFAM) 0 1 2 3 4 5 Homo sapiens 0.24 DNA-directed RNA polymerase, mitochondrial 1 1 Homo sapiens DNA (56-mer) 1 2 3 Homo sapiens Dimethyladenosine transferase 2, mitochondrial 1 4 Homo sapiens DNA-directed RNA polymerase, mitochondrial Homo sapiens 0.13463234400000001 1 Escherichia coli BL21 LEVO SASPQEQDQDRRKDWGHVELLEVLQARVRQLQAESVSEVVVNRVDVARLPECGSGDGSLQPPRKVQMGAKDATPVPCGRW AKILEKDKRTQQMRMQRLKAKLQMPFQSGEFKALTRRLQVEPRLLSKQMAGCLEDCTRQAPESPWEEQLARLLQEAPGKL SLDVEQAPSGQHSQAQLSGQQQRLLAFFKCCLLTDQLPLAHHLLVVHHGQRQKRKLLTLDMYNAVMLGWARQGAFKELVY VLFMVKDAGLTPDLLSYAAALQCMGRQDQDAGTIERCLEQMSQEGLKLQALFTAVLLSEEDRATVLKAVHKVKPTFSLPP QLPPPVNTSKLLRDVYAKDGRVSYPKLHLPLKTLQCLFEKQLHMELASRVCVVSVEKPTLPSKEVKHARKTLKTLRDQWE KALCRALRETKNRLEREVYEGRFSLYPFLCLLDEREVVRMLLQVLQALPAQGESFTTLARELSARTFSRHVVQRQRVSGQ VQALQNHYRKYLCLLASDAEVPEPCLPRQYWEELGAPEALREQPWPLPVQMELGKLLAEMLVQATQMPCSLDKPHRSSRL VPVLYHVYSFRNVQQIGILKPHPAYVQLLEKAAEPTLTFEAVDVPMLCPPLPWTSPHSGAFLLSPTKLMRTVEGATQHQE LLETCPPTALHGALDALTQLGNCAWRVNGRVLDLVLQLFQAKGCPQLGVPAPPSEAPQPPEAHLPHSAAPARKAELRREL AHCQKVAREMHSLRAEALYRLSLAQHLRDRVFWLPHNMDFRGRTYPCPPHFNHLGSDVARALLEFAQGRPLGPHGLDWLK IHLVNLTGLKKREPLRKRLAFAEEVMDDILDSADQPLTGRKWWMGAEEPWQTLACCMEVANAVRASDPAAYVSHLPVHQD GSCNGLQHYAALGRDSVGAASVNLEPSDVPQDVYSGVAAQVEVFRRQDAQRGMRVAQVLEGFITRKVVKQTVMTVVYGVT RYGGRLQIEKRLRELSDFPQEFVWEASHYLVRQVFKSLQEMFSGTRAIQHWLTESARLISHMGSVVEWVTPLGVPVIQPY RLDSKVKQIGGGIQSITYTHNGDISRKPNTRKQKNGFPPNFIHSLDSSHMMLTALHCYRKGLTFVSVHDCYWTHAADVSV MNQVCREQFVRLHSEPILQDLSRFLVKRFCSEPQKILEASQLKETLQAVPKPGAFDLEQVKRSTYFFS O00411 2.7.7.6 Non-template strand DNA (56-mer) synthetic construct 0.019652648 1 (DA)(DT)(DG)(DT)(DG)(DT)(DT)(DA)(DG)(DT)(DT)(DG)(DG)(DG)(DG)(DG)(DG)(DT)(DG)(DA) (DC)(DT)(DG)(DT)(DT)(DA)(DA)(DA)(DA)(DG)(DT)(DG)(DC)(DA)(DT)(DA)(DC)(DC)(DG)(DA) (DA)(DC)(DA)(DA)(DA)(DG)(DA)(DT)(DA)(DA)(DA)(DA)(DT)(DT)(DT)(DG)(DA)(DA)(DA)(DT) (DC)(DT)(DG) DNA Template strand DNA (56-MER) synthetic construct 0.018867131 1 (DC)(DA)(DG)(DT)(DT)(DT)(DC)(DA)(DA)(DA)(DT)(DT)(DT)(DT)(DA)(DT)(DC)(DT)(DC)(DC) (DA)(DG)(DG)(DC)(DG)(DG)(DT)(DA)(DT)(DG)(DC)(DA)(DC)(DT)(DT)(DT)(DT)(DA)(DA)(DC) (DA)(DG)(DT)(DC)(DA)(DC)(DC)(DC)(DC)(DC)(DC)(DA)(DA)(DC)(DT)(DA)(DA)(DC)(DA)(DC) (DA)(DT) DNA Dimethyladenosine transferase 2, mitochondrial Homo sapiens 0.038901309 1 Escherichia coli B LEVO PPRKASKASLDFKRYVTDRRLAETLAQIYLGKPSRPPHLLLECNPGPGILTQALLEAGAKVVALESDKTFIPHLESLGKN LDGKLRVIHCDFFKLDPRSGGVIKPPAMSSRGLFKNLGIEAVPWTADIPLKVVGMFPSRGEKRALWKLAYDLYSCTSIYK FGRIEVNMFIGEKEFQKLMADPGNPDLYHVLSVIWQLACEIKVLHMEPWSSFDIYTRKGPLENPKRRELLDQLQQKLYLI QMIPRQNLFTKNLTPMNYNIFFHLLKHCFGRRSATVIDHLRSLTPLDARDILMQIGKQEDEKVVNMHPQDFKTLFETIER SKDCAYKWLYDETLEDR Q9H5Q4 2.1.1.- RNA (5'-D(*(GTP))-R(P*A)-3') synthetic construct 0.000789413 1 (GTP)A GUANOSINE-5'-TRIPHOSPHATE 0.0005231799999999999 1 GTP MAGNESIUM ION 2.4305e-05 1 MG singleParticle particle 1.2 8.0

30 mM Tris-HCl pH 8, 200 mM NaCl, 3 mM DTT, 10 mM MgCl2

Quantifoil R1.2/1.3 GOLD 300 CARBON HOLEY GLOW DISCHARGE 45 AIR 0.03 ETHANE 100 277.15 FEI VITROBOT MARK IV

incubated on the grid for 5 seconds prior to double sided blotting for 3.5 seconds at force 1.

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