2019-03-06RELRCSBRCSBRCSB2018-12-052019-02-202019-03-062019-03-06Structure of bacteriophage T7 lagging-strand DNA polymerase (D5A/E7A)/Trx interacting with primase domains of two gp4 subunits (E and F), with gp4 helicase bound to a DNA fork and dTTP (LagL1)Gao YFox TVal NYang WGao YCui YFox TLin SWang Hde Val NZhou ZHYang WStructures and operating principles of the replisome.ScienceUS363201930679383doi:10.1126/science.aav70031095-92030038SCIEASEMD-0387associated EM volumeEMD-0364other EM volumeEMD-0365other EM volumeEMD-0379other EM volumeEMD-0380other EM volumeEMD-0381other EM volumeEMD-0382other EM volumeEMD-0386other EM volumeEMD-0388other EM volumeEMD-0389other EM volumeEMD-0390other EM volumeEMD-0391other EM volumeEMD-0392other EM volumeEMD-0393other EM volumeEMD-0394other EM volumeEMD-0395other EM volumeT7 DNA polymerase/Trx interacting with two primase domains (E and F) of T7 helicase (gp4) in complex with a fork DNA substrate and dTTP (from gp4-gp5 lagging-strand complex LagL1)T7 DNA polymerase/Trx interacting with two primase domains (E and F) of T7 helicase (gp4) in complex with a fork DNA substrate and dTTP (from gp4-gp5 lagging-strand complex LagL1)012345Enterobacteria phage T7Escherichia coligene product 4 helicase-primaseEnterobacteria phage T7LEVOMDNSHDSDSVFLYHIPCDNCGSSDGNSLFSDGHTFCYVCEKWTAGNEDTKERASKRKPSG
GKPMTYNVWNFGESNGRYSALTARGISKETCQKAGYWIAKVDGVMYQVADYRDQNGNIVS
QKVRDKDKNFKTTGSHKSDALFGKHLWNGGKKIVVTEGEIDMLTVMELQDCKYPVVSLGH
GASAAKKTCAANYEYFDQFEQIILMFDMDEAGRKAVEEAAQVLPAGKVRVAVLPCKDANE
CHLNGHDREIMEQVWNAGPWIPDGVVSALSLRERIREHLSSEESVGLLFSGCTGINDKTL
GARGGEVIMVTSGSGMGKSTFVRQQALQWGTAMGKKVGLAMLQESVEETAEDLIGLHNRV
RLRQSDSLKREIIENGKFDQWFDELFGNDTFHLYDSFAEAETDRLLAKLAYMRSGLGCDV
IILDHISIVVSASGESDERKMIDNLMTKLKGFAKSTGVVLVVICHLKNPDKGKAHEEGRP
VSITDLRGSGALRQLSDTIIALERNQQGDMPNLVLVRILKCRFTGDTGIAGYMEYNKETG
WLEPSSYSGEEESHSESTDWSNDTDFgene product 5 DNA polymeraseEnterobacteria phage T7LEVOMIVSAIAANALLESVTKFHCGVIYDYSTAEYVSYRPSDFGAYLDALEAEVARGGLIVFHN
GHKYDVPALTKLAKLQLNREFHLPRENCIDTLVLSRLIHSNLKDTDMGLLRSGKLPGKRF
GSHALEAWGYRLGEMKGEYKDDFKRMLEEQGEEYVDGMEWWNFNEEMMDYNVQDVVVTKA
LLEKLLSDKHYFPPEIDFTDVGYTTFWSESLEAVDIEHRAAWLLAKQERNGFPFDTKAIE
ELYVELAARRSELLRKLTETFGSWYQPKGGTEMFCHPRTGKPLPKYPRIKTPKVGGIFKK
PKNKAQREGREPCELDTREYVAGAPYTPVEHVVFNPSSRDHIQKKLQEAGWVPTKYTDKG
APVVDDEVLEGVRVDDPEKQAAIDLIKEYLMIQKRIGQSAEGDKAWLRYVAEDGKIHGSV
NPNGAVTGRATHAFPNLAQIPGVRSPYGEQCRAAFGAEHHLDGITGKPWVQAGIDASGLE
LRCLAHFMARFDNGEYAHEILNGDIHTKNQIAAELPTRDNAKTFIYGFLYGAGDEKIGQI
VGAGKERGKELKKKFLENTPAIAALRESIQQTLVESSQWVAGEQQVKWKRRWIKGLDGRK
VHVRSPHAALNTLLQSAGALICKLWIIKTEEMLVEKGLKHGWDGDFAYMAWVHDEIQVGC
RTEEIAQVVIETAQEAMRWVGDHWNFRCLLDTEGKMGPNWAICHThioredoxinEscherichia coliLEVOMSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEIADEYQGKLTVAKLN
IDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLDANLATemplateEnterobacteria phage T7(DT)(DT)(DT)(DG)(DG)(DT)(DC)(DA)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DT)(DA)(DC)(DG)(DG)(DA)(DG)(DT)(DC)(DG)(DT)(DT)(DT)(DC)(DG)(DA)(DC)(DT)(DC)(DC)(DG)(DT)(DT)(DA)(DT)(DC)(DA)(DC)(DG)(DC)(DT)(DA)(DT)(DG)(DT)(DC)(DG)(DT)(DC)(DA)(DA)(DG)(DT)(DT)(DG)(DT)(DA)(DC)(DC)DNAPrimerEnterobacteria phage T7(DG)(DG)(DT)(DA)(DC)(DA)(DA)(DC)(DT)(DT)(DG)(DA)(DC)(DG)(DA)(DC)(DA)(DT)(DA)(DG)(DC)(DG)(DT)(DG)(DOA)DNAsingleParticleparticle7.550.0Tris150.0KClpotassium chloride3.0DTTunspecifiedETHANE100295FEI VITROBOT MARK IFEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN300GATAN K2 SUMMIT (4k x 4k)COUNTING40.011T8E1T8E, 1NUIC16.6FSC 0.143 CUT-OFFRELION30550MAXIMUM LIKELIHOODRELIONMAXIMUM LIKELIHOODRELION1T8EFLEXIBLE FITREAL