IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Protein Science 2021 imex curation Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". lp212@cam.ac.uk Only protein-protein interactions imex curation in vitro In vitro rna dna pol assay rna directed dna polymerase assay rna dna pol assay tag fluorescence tag visualisation by fluorescence tag fluorescence Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". lp212@cam.ac.uk Accepted 2021-DEC-20 AT 10:43 GMT AT 10:43 GMT by KALPANAP Protein Science 2021 Only protein-protein interactions imex curation imex curation Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Protein Science 2021 imex curation Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". lp212@cam.ac.uk Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight by staining molecular weight estimation by staining weight by staining Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". lp212@cam.ac.uk Accepted 2021-DEC-20 AT 10:43 GMT AT 10:43 GMT by KALPANAP Protein Science 2021 Only protein-protein interactions imex curation imex curation Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Structural basis for the interaction of SARS-CoV-2 virulence factor nsp1 with DNA polymerase a–primase Protein Science 2021 imex curation Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". lp212@cam.ac.uk Only protein-protein interactions imex curation in vitro In vitro 3d-em 3D electron microscopy 3D-EM electron tomog structure determination identification by structure determination identification by structure determination structure determination Kilkenny M, Kilkenny M Virus - Publications including interactions involving viral proteins pdbe strep tag sequence does not correspond to stated "double StrepII tag" - so just entered as "strep tag". Samples were cross linked with BS3 ((bis(sulfosuccinimidyl)suberate)) before freezing. lp212@cam.ac.uk Accepted 2021-DEC-20 AT 10:43 GMT AT 10:43 GMT by KALPANAP Protein Science 2021 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method pri1_human DNA primase small subunit DNA primase 49 kDa subunit PRIM1 protein protein human Homo sapiens Human METFDPTELPELLKLYYRRLFPYSQYYRWLNYGGVIKNYFQHREFSFTLKDDIYIRYQSFNNQSDLEKEMQKMNPYKIDIGAVYSHRPNQHNTVKLGAFQAQEKELVFDIDMTDYDDVRRCCSSADICPKCWTLMTMAIRIIDRALKEDFGFKHRLWVYSGRRGVHCWVCDESVRKLSSAVRSGIVEYLSLVKGGQDVKKKVHLSEKIHPFIRKSINIIKKYFEEYALVNQDILENKESWDKILALVPETIHDELQQSFQKSHNSLQRWEHLKKVASRYQNNIKNDKYGPWLEWEIMLQYCFPRLDINVSKGINHLLKSPFSVHPKTGRISVPIDLQKVDQFDPFTVPTISFICRELDAISTNEEEKEENEAESDVKHRTRDYKKTSLAPYVKVFEHFLENLDKSRKGELLKKSDLQKDF 9B5AC900E0C3CCE8 dna45 ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TTCTTTATCTCTTATTTCCTTCTATTTTCCACGCGCCTTCTATTT tetra-mu3-sulfido-tetrairon tetra-mu(3)-sulfido-tetrairon (IRON/SULFUR CLUSTER) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dpola_human DNA polymerase alpha catalytic subunit POLA1 POLA DNA polymerase alpha catalytic subunit p180 protein protein human Homo sapiens Human MAPVHGDDSLSDSGSFVSSRARREKKSKKGRQEALERLKKAKAGEKYKYEVEDFTGVYEEVDEEQYSKLVQARQDDDWIVDDDGIGYVEDGREIFDDDLEDDALDADEKGKDGKARNKDKRNVKKLAVTKPNNIKSMFIACAGKKTADKAVDLSKDGLLGDILQDLNTETPQITPPPVMILKKKRSIGASPNPFSVHTATAVPSGKIASPVSRKEPPLTPVPLKRAEFAGDDVQVESTEEEQESGAMEFEDGDFDEPMEVEEVDLEPMAAKAWDKESEPAEEVKQEADSGKGTVSYLGSFLPDVSCWDIDQEGDSSFSVQEVQVDSSHLPLVKGADEEQVFHFYWLDAYEDQYNQPGVVFLFGKVWIESAETHVSCCVMVKNIERTLYFLPREMKIDLNTGKETGTPISMKDVYEEFDEKIATKYKIMKFKSKPVEKNYAFEIPDVPEKSEYLEVKYSAEMPQLPQDLKGETFSHVFGTNTSSLELFLMNRKIKGPCWLEVKSPQLLNQPVSWCKVEAMALKPDLVNVIKDVSPPPLVVMAFSMKTMQNAKNHQNEIIAMAALVHHSFALDKAAPKPPFQSHFCVVSKPKDCIFPYAFKEVIEKKNVKVEVAATERTLLGFFLAKVHKIDPDIIVGHNIYGFELEVLLQRINVCKAPHWSKIGRLKRSNMPKLGGRSGFGERNATCGRMICDVEISAKELIRCKSYHLSELVQQILKTERVVIPMENIQNMYSESSQLLYLLEHTWKDAKFILQIMCELNVLPLALQITNIAGNIMSRTLMGGRSERNEFLLLHAFYENNYIVPDKQIFRKPQQKLGDEDEEIDGDTNKYKKGRKKAAYAGGLVLDPKVGFYDKFILLLDFNSLYPSIIQEFNICFTTVQRVASEAQKVTEDGEQEQIPELPDPSLEMGILPREIRKLVERRKQVKQLMKQQDLNPDLILQYDIRQKALKLTANSMYGCLGFSYSRFYAKPLAALVTYKGREILMHTKEMVQKMNLEVIYGDTDSIMINTNSTNLEEVFKLGNKVKSEVNKLYKLLEIDIDGVFKSLLLLKKKKYAALVVEPTSDGNYVTKQELKGLDIVRRDWCDLAKDTGNFVIGQILSDQSRDTIVENIQKRLIEIGENVLNGSVPVSQFEINKALTKDPQDYPDKKSLPHVHVALWINSQGGRKVKAGDTVSYVICQDGSNLTASQRAYAPEQLQKQDNLTIDTQYYLAQQIHPVVARICEPIDGIDAVLIATWLGLDPTQFRVHHYHKDEENDALLGGPAQLTDEEKYRDCERFKCPCPTCGTENIYDNVFDGSGTDMEPSLYRCSNIDCKASPLTFTVQLSNKLIMDIRRFIKKYYDGWLICEEPTCRNRTRHLPLQFSRTGPLCPACMKATLQPEYSDKSLYTQLCFYRYIFDAECALEKLTTDHEKDKLKKQFFTPKVLQDYRKLKNTAEQFLSRSGYSEVNLSKLFAGCAVKS DC40C3EDD6F4B495 rna13 rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) GGCGCGUGGAAAA p0c6u8-pro_0000338255 Host translation inhibitor nsp1 ORF1a polyprotein 1a protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus MESLVLGVNEKTHVQLSLPVLQVRDVLVRGFGDSVEEALSEAREHLKNGTCGLVELEKGVLPQLEQPYVFIKRSDALSTNHGHKVVELVAEMDGIQYGRSGITLGVLVPHVGETPIAYRNVLLRKNGNKGAGGHSYGIDLKSYDLGDELGTDPIEDYEQNWNTKHGSGALRELTRELNGG 1 180 B76E7770809587A7 pri2_human DNA primase large subunit DNA primase 58 kDa subunit PRIM2 PRIM2A protein protein human Homo sapiens Human MEFSGRKWRKLRLAGDQRNASYPHCLQFYLQPPSENISLIEFENLAIDRVKLLKSVENLGVSYVKGTEQYQSKLESELRKLKFSYRENLEDEYEPRRRDHISHFILRLAYCQSEELRRWFIQQEMDLLRFRFSILPKDKIQDFLKDSQLQFEAISDEEKTLREQEIVASSPSLSGLKLGFESIYKIPFADALDLFRGRKVYLEDGFAYVPLKDIVAIILNEFRAKLSKALALTARSLPAVQSDERLQPLLNHLSHSYTGQDYSTQGNVGKISLDQIDLLSTKSFPPCMRQLHKALRENHHLRHGGRMQYGLFLKGIGLTLEQALQFWKQEFIKGKMDPDKFDKGYSYNIRHSFGKEGKRTDYTPFSCLKIILSNPPSQGDYHGCPFRHSDPELLKQKLQSYKISPGGISQILDLVKGTHYQVACQKYFEMIHNVDDCGFSLNHPNQFFCESQRILNGGKDIKKEPIQPETPQPKPSVQKTKDASSALASLNSSLEMDMEGLEDYFSEDS 80195D48103F87A7 dpoa2_human DNA polymerase alpha subunit B DNA polymerase alpha 70 kDa subunit POLA2 protein protein human Homo sapiens Human MSASAQQLAEELQIFGLDCEEALIEKLVELCVQYGQNEEGMVGELIAFCTSTHKVGLTSEILNSFEHEFLSKRLSKARHSTCKDSGHAGARDIVSIQELIEVEEEEEILLNSYTTPSKGSQKRAISTPETPLTKRSVSTRSPHQLLSPSSFSPSATPSQKYNSRSNRGEVVTSFGLAQGVSWSGRGGAGNISLKVLGCPEALTGSYKSMFQKLPDIREVLTCKIEELGSELKEHYKIEAFTPLLAPAQEPVTLLGQIGCDSNGKLNNKSVILEGDREHSSGAQIPVDLSELKEYSLFPGQVVIMEGINTTGRKLVATKLYEGVPLPFYQPTEEDADFEQSMVLVACGPYTTSDSITYDPLLDLIAVINHDRPDVCILFGPFLDAKHEQVENCLLTSPFEDIFKQCLRTIIEGTRSSGSHLVFVPSLRDVHHEPVYPQPPFSYSDLSREDKKQVQFVSEPCSLSINGVIFGLTSTDLLFHLGAEEISSSSGTSDRFSRILKHILTQRSYYPLYPPQEDMAIDYESFYVYAQLPVTPDVLIIPSELRYFVKDVLGCVCVNPGRLTKGQVGGTFARLYLRRPAADGAERQSPCIAVQVVRI F5CBD7E022ADDDBF pol_prim_human-pro_0000338255-1 3 10 unspecified role unspecified role neutral component neutral component purified purified middle domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 19 n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 21 n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 19 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 21 n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction Supp. Table 1 pol_prim_human-pro_0000338255-2 2 10 unspecified role unspecified role bait bait purified purified P0C6U8-PRO_0000338255:p.Leu27Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain L D Thu Jan 06 19:11:04 GMT 2022 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu27Asp' n-terminus mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined middle domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0C6U8-PRO_0000338255:p.Val28Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V D Thu Jan 06 19:50:52 GMT 2022 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val28Asp' P0C6U8-PRO_0000338255:p.Val111Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V D Thu Jan 06 20:22:21 GMT 2022 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val111Asp' ecoli Escherichia coli (strain K12) 17 unspecified role unspecified role prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction Figure 1,2 Both full-length and truncated forms of purified primosome were pulled-down. pro_0000338255-pol_prim_human-1 1 17 enzyme enzyme neutral component neutral component purified purified n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus strep tag strep tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 9 enzyme target enzyme target substrate substrate neutral component neutral component whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5'-prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 6 ancillary ancillary neutral component neutral component whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 49 46 direct interaction direct interaction Figure 1,2 NSP1s had no effect. pol-prim_human DNA polymerase alpha:primase complex DPOLA-DPOA2-PRI1-PRI2 complex POLA1:POLA2:PRIM1:PRIM2 DNA polymerase alpha:primase complex Alpha DNA polymerase:primase complex heterotetrameric polymerase alpha holoenzyme pol-prim primosome Pol alpha-primase complex polymerase alpha-primase complex 7 cofactor cofactor coenzyme pola1 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prim2 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 8 enzyme enzyme pola2 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain prim2 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 4fe-4s binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 12 unspecified role unspecified role pola1 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 11 enzyme enzyme pola1 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain prim1 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 55 51 59 52 56 53 54 58 physical association physical association human Homo sapiens Human stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction Initiates DNA replication by synthesizing short RNA primers on the leading and lagging strand templates in a minimum of five steps: template binding, NTP binding, dinucleotide formation, extension to a functional RNA primer, and primer transfer to the POLA catalytic site for elongation into hybrid primers of about 35 nucleotides. Heterotetramer created with intact-jami-dbscripts