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/ http://www.ebi.ac.uk/ chilkova-2003-1 The quaternary structure of DNA polymerase epsilon from Saccharomyces cerevisiae. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Chilkova O., Jonsson BH., Johansson E. The POL2 gene was subcloned into one plasmid and DPB2, DPB3, and DPB4 were subcloned into the second plasmid. All genes were cloned under the control of the galactose-inducible GAL1-10 promoter. The complex was purified by successive phosphocellulose, MonoQ, and MonoS chromatography, and the enzyme was finally purified to homogeneity by Superose 6 gel filtration column. The proteins were visualized by colloidal Coomassie staining. 1 & 3 erik.johansson@medchem.umu.se J. Biol. Chem. (0021-9258) 2003 IMEx Chilkova O., Jonsson BH., Johansson E., Chilkova O., Jonsson BH., Johansson E. Only protein-protein interactions imex curation imex curation chilkova-2003-2 The quaternary structure of DNA polymerase epsilon from Saccharomyces cerevisiae. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast density sedimentation cosedimentation through density gradient density sedimentation predetermined predetermined participant predetermined Chilkova O., Jonsson BH., Johansson E. The POL2 gene was subcloned into one plasmid and DPB2, DPB3, and DPB4 were subcloned into the second plasmid. All genes were cloned under the control of the galactose-inducible GAL1-10 promoter. The complex was purified by successive phosphocellulose, MonoQ, and MonoS chromatography, and the enzyme was finally purified to homogeneity by Superose 6 gel filtration column. The proteins were visualized by colloidal Coomassie staining. 1 & 3 erik.johansson@medchem.umu.se J. Biol. Chem. (0021-9258) 2003 IMEx Chilkova O., Jonsson BH., Johansson E., Chilkova O., Jonsson BH., Johansson E. Only protein-protein interactions imex curation imex curation chilkova-2003-3 The quaternary structure of DNA polymerase epsilon from Saccharomyces cerevisiae. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast cosedimentation cosedimentation predetermined predetermined participant predetermined Chilkova O., Jonsson BH., Johansson E. The POL2 gene was subcloned into one plasmid and DPB2, DPB3, and DPB4 were subcloned into the second plasmid. All genes were cloned under the control of the galactose-inducible GAL1-10 promoter. The complex was purified by successive phosphocellulose, MonoQ, and MonoS chromatography, and the enzyme was finally purified to homogeneity by Superose 6 gel filtration column. The proteins were visualized by colloidal Coomassie staining. 1 & 3 erik.johansson@medchem.umu.se J. Biol. Chem. (0021-9258) 2003 IMEx Chilkova O., Jonsson BH., Johansson E., Chilkova O., Jonsson BH., Johansson E. Only protein-protein interactions imex curation imex curation dpb4_yeast DNA polymerase epsilon subunit D DPB4 DNA polymerase II subunit D YDR121W protein protein yeast Saccharomyces cerevisiae Baker's yeast MPPKGWRKDAQGNYPTTSYIKEQENITIQDLLFPKSTIVNLAREVPQQSGKKLLINKDASLALQRGATVFVNHLLLFAREIAKSQDKKSCSVDDVLSALDHIGHSALKGPVRDKLDEYQAAVEQRKKEKLDSGEVDADGDIDMGEDKENVPVEKVKEHDEIEEQGDALQDVEESSEKKQKTESQDVETRVQNLEQT dpb2_yeast DNA polymerase epsilon subunit B DPB2 YPR175W P9705.7 DNA polymerase II subunit 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFGSGNVLPVKIQPPLLRPLAYRVLSRKYGLSIKSDGLSALAEFVGTNIGANWRQGPATIKFLEQFAAVWKQQERGLFIDQSGVKEVIQEMKEREKVEWSHEHPIQHEENILGRTDDDENNSDDEMPIAADSSLQNVSLSSPMRQPTERDEYKQPFKPESSKALDWRDYFKVINASQQQRFSYNPHKMQFIFVPNKKQNGLGGIAGFLPDIEDKVQMFLTRYYLTNDRVMRNENFQNSDMFNPLSSMVSLQNELSNTNRQQQSSSMSITPIKNLLGRDAQNFLLLGLLNKNFKGNWSLEDPSGSVEIDISQTIPTQGHYYVPGCMVLVEGIYYSVGNKFHVTSMTLPPGERREITLETIGNLDLLGIHGISNNNFIARLDKDLKIRLHLLEKELTDHKFVILGANLFLDDLKIMTALSKILQKLNDDPPTLLIWQGSFTSVPVFASMSSRNISSSTQFKNNFDALATLLSRFDNLTENTTMIFIPGPNDLWGSMVSLGASGTLPQDPIPSAFTKKINKVCKNVVWSSNPTRIAYLSQEIVIFRDDLSGRFKRHRLEFPFNESEDVYTENDNMMSKDTDIVPIDELVKEPDQLPQKVQETRKLVKTILDQGHLSPFLDSLRPISWDLDHTLTLCPIPSTMVLCDTTSAQFDLTYNGCKVINPGSFIHNRRARYMEYVPSSKKTIQEEIYI 0936E3FC90552788 dpb3_yeast DNA polymerase epsilon subunit C DPB3 YBR278W YBR2015 DNA polymerase II subunit C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNLVKEKAPVFPISKVKKIAKCDPEYVITSNVAISATAFAAELFVQNLVEESLVLAQLNSKGKTSLRLSLNSIEECVEKRDNFRFLEDAIKQLKKNSALDKKRELNMQPGRSDQEVVIEEPELHEDDGVEEEEEEDEVSEEEEPVHNEELLDDSKDQQNDKSTRSVASLLSRFQYKSALDVGEHSDSSDIEVDHTKSTDP 288913917D49C7F9 dpoe_yeast DNA polymerase epsilon catalytic subunit A POL2 DUN2 YNL262W N0825 DNA polymerase II subunit A 3'-5' exodeoxyribonuclease protein protein yeast Saccharomyces cerevisiae Baker's yeast MMFGKKKNNGGSSTARYSAGNKYNTLSNNYALSAQQLLNASKIDDIDSMMGFERYVPPQYNGRFDAKDIDQIPGRVGWLTNMHATLVSQETLSSGSNGGGNSNDGERVTTNQGISGVDFYFLDEEGGSFKSTVVYDPYFFIACNDESRVNDVEELVKKYLESCLKSLQIIRKEDLTMDNHLLGLQKTLIKLSFVNSNQLFEARKLLRPILQDNANNNVQRNIYNVAANGSEKVDAKHLIEDIREYDVPYHVRVSIDKDIRVGKWYKVTQQGFIEDTRKIAFADPVVMAFDIETTKPPLKFPDSAVDQIMMISYMIDGEGFLITNREIISEDIEDFEYTPKPEYPGFFTIFNENDEVALLQRFFEHIRDVRPTVISTFNGDFFDWPFIHNRSKIHGLDMFDEIGFAPDAEGEYKSSYCSHMDCFRWVKRDSYLPQGSQGLKAVTQSKLGYNPIELDPELMTPYAFEKPQHLSEYSVSDAVATYYLYMKYVHPFIFSLCTIIPLNPDETLRKGTGTLCEMLLMVQAYQHNILLPNKHTDPIERFYDGHLLESETYVGGHVESLEAGVFRSDLKNEFKIDPSAIDELLQELPEALKFSVEVENKSSVDKVTNFEEIKNQITQKLLELKENNIRNELPLIYHVDVASMYPNIMTTNRLQPDSIKAERDCASCDFNRPGKTCARKLKWAWRGEFFPSKMDEYNMIKRALQNETFPNKNKFSKKKVLTFDELSYADQVIHIKKRLTEYSRKVYHRVKVSEIVEREAIVCQRENPFYVDTVKSFRDRRYEFKGLAKTWKGNLSKIDPSDKHARDEAKKMIVLYDSLQLAHKVILNSFYGYVMRKGSRWYSMEMAGITCLTGATIIQMARALVERVGRPLELDTDGIWCILPKSFPETYFFTLENGKKLYLSYPCSMLNYRVHQKFTNHQYQELKDPLNYIYETHSENTIFFEVDGPYKAMILPSSKEEGKGIKKRYAVFNEDGSLAELKGFELKRRGELQLIKNFQSDIFKVFLEGDTLEGCYSAVASVCNRWLDVLDSHGLMLEDEDLVSLICENRSMSKTLKEYEGQKSTSITTARRLGDFLGEDMVKDKGLQCKYIISSKPFNAPVTERAIPVAIFSADIPIKRSFLRRWTLDPSLEDLDIRTIIDWGYYRERLGSAIQKIITIPAALQGVSNPVPRVEHPDWLKRKIATKEDKFKQTSLTKFFSKTKNVPTMGKIKDIEDLFEPTVEEDNAKIKIARTTKKKAVSKRKRNQLTNEEDPLVLPSEIPSMDEDYVGWLNYQKIKWKIQARDRKRRDQLFGNTNSSRERSALGSMIRKQAESYANSTWEVLQYKDSGEPGVLEVFVTINGKVQNITFHIPKTIYMKFKSQTMPLQKIKNCLIEKSSASLPNNPKTSNPAGGQLFKITLPESVFLEEKENCTSIFNDENVLGVFEGTITPHQRAIMDLGASVTFRSKAMGALGKGIQQGFEMKDLSMAENERYLSGFSMDIGYLLHFPTSIGYEFFSLFKSWGDTITILVLKPSNQAQEINASSLGQIYKQMFEKKKGKIETYSYLVDIKEDINFEFVYFTDISKLYRRLSQETTKLKEERGLQFLLLLQSPFITKLLGTIRLLNQMPIVKLSLNEVLLPQLNWQPTLLKKLVNHVLSSGSWISHLIKLSQYSNIPICNLRLDSMDYIIDVLYARKLKKENIVLWWNEKAPLPDHGGIQNDFDLNTSWIMNDSEFPKINNSGVYDNVVLDVGVDNLTVNTILTSALINDAEGSDLVNNNMGIDDKDAVINSPSEFVHDAFSNDALNVLRGMLKEWWDEALKENSTADLLVNSLASWVQNPNAKLFDGLLRYHVHNLTKKALLQLVNEFSALGSTIVYADRNQILIKTNKYSPENCYAYSQYMMKAVRTNPMFSYLDLNIKRYWDLLIWMDKFNFSGLACIEIEEKENQDYTAVSQWQLKKFLSPIYQPEFEDWMMIILDSMLKTKQSYLKLNSGTQRPTQIVNVKKQDKEDSVENSLNGFSHLFSKPLMKRVKKLFKNQQEFILDPQYEADYVIPVLPGSHLNVKNPLLELVKSLCHVMLLSKSTILEIRTLRKELLKIFELREFAKVAEFKDPSLSLVVPDFLCEYCFFISDIDFCKAAPESIFSCVRCHKAFNQVLLQEHLIQKLRSDIESYLIQDLRCSRCHKVKRDYMSAHCPCAGAWEGTLPRESIVQKLNVFKQVAKYYGFDILLSCIADLTI CBCDDE2AB147D65B pol2-dpb2-1 Isolation of the DNA polymerase epsilon complex by gel filtration 332574 N/A 332577 unspecified role unspecified role neutral component neutral component N/A 332578 unspecified role unspecified role neutral component neutral component N/A 332579 unspecified role unspecified role neutral component neutral component N/A 332580 unspecified role unspecified role neutral component neutral component physical association physical association false false false Stokes radius of the complex was estimated to be 74.5A pol2-dpb2-2 Isolation of the DNA polymerase epsilon complex by glycerol gradient centrifugation 332575 N/A 332577 unspecified role unspecified role neutral component neutral component N/A 332578 unspecified role unspecified role neutral component neutral component N/A 332579 unspecified role unspecified role neutral component neutral component N/A 332580 unspecified role unspecified role neutral component neutral component physical association physical association false false false The sedimentation coefficient for Pol epsilon was determined to be 11.9 S. Calculated molecular mass = 371 kDa. pol2-dpb2-3 Isolation of the DNA polymerase epsilon complex by sedimentation equilibrium ultracentrifugation 332576 N/A 332577 unspecified role unspecified role neutral component neutral component N/A 332578 unspecified role unspecified role neutral component neutral component N/A 332579 unspecified role unspecified role neutral component neutral component N/A 332580 unspecified role unspecified role neutral component neutral component physical association physical association false false false The molecular mass of Pol epsilon w= 366 kDa under these conditions. A comparison with the predicted distribution of a monomeric versus a dimeric assembly showed that Pol epsilon was present as a monomer in the solution under all conditions investigated