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/ The yeast capping enzyme represses RNA polymerase II transcription. The yeast capping enzyme represses RNA polymerase II transcription. Mol. Cell (1097-2765) 2002 imex curation Myers LC., Lacomis L., Erdjument-Bromage H., Tempst P. Complex isolated by anion exchange, hydroxylapatite (HAP), and hydrophobic-chromatography, followed by final resolution on a Mono-Q column. larry.myeers@dartmouth.edu 23-MAY-2007: Contacted by JYOTI. Returned mail. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Myers LC., Lacomis L., Erdjument-Bromage H., Tempst P. Revisit when immunodepletion added to methods Participants identified by immunodepletion in that a DE400 fraction was incubated with anti-Cet1p antibodies immobilized on protein-A beads, and the supernatant was recovered. An equal volume of load (DE400) and supernatant (DE400[minusCet1/Ceg1]) was analyzed by Western blotting. larry.myeers@dartmouth.edu Accepted 2007-JUN-8 by DAVET. 23-MAY-2007: Contacted by JYOTI. Returned mail. Mol. Cell (1097-2765) 2002 Only protein-protein interactions imex curation The yeast capping enzyme represses RNA polymerase II transcription. The yeast capping enzyme represses RNA polymerase II transcription. Mol. Cell (1097-2765) 2002 imex curation Myers LC., Lacomis L., Erdjument-Bromage H., Tempst P. Complex isolated by anion exchange, hydroxylapatite (HAP), and hydrophobic-chromatography, followed by final resolution on a Mono-Q column. larry.myeers@dartmouth.edu 23-MAY-2007: Contacted by JYOTI. Returned mail. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast chromatography chromatography technology column chromatography chromatography fingerprinting peptide massfingerprinting fingerprinting Myers LC., Lacomis L., Erdjument-Bromage H., Tempst P. Complex isolated by anion exchange, hydroxylapatite (HAP), and hydrophobic-chromatography, followed by final resolution on a Mono-Q column. larry.myeers@dartmouth.edu Accepted 2007-JUN-8 by DAVET. 23-MAY-2007: Contacted by JYOTI. Returned mail. Mol. Cell (1097-2765) 2002 Only protein-protein interactions imex curation mce1_yeast mRNA-capping enzyme subunit alpha CEG1 YGL130W G2853 mRNA guanylyltransferase GTP--RNA guanylyltransferase protein protein yeast Saccharomyces cerevisiae Baker's yeast MVLAMESRVAPEIPGLIQPGNVTQDLKMMVCKLLNSPKPTKTFPGSQPVSFQHSDVEEKLLAHDYYVCEKTDGLRVLMFIVINPVTGEQGCFMIDRENNYYLVNGFRFPRLPQKKKEELLETLQDGTLLDGELVIQTNPMTKLQELRYLMFDCLAINGRCLTQSPTSSRLAHLGKEFFKPYFDLRAAYPNRCTTFPFKISMKHMDFSYQLVKVAKSLDKLPHLSDGLIFTPVKAPYTAGGKDSLLLKWKPEQENTVDFKLILDIPMVEDPSLPKDDRNRWYYNYDVKPVFSLYVWQGGADVNSRLKHFDQPFDRKEFEILERTYRKFAELSVSDEEWQNLKNLEQPLNGRIVECAKNQETGAWEMLRFRDDKLNGNHTSVVQKVLESINDSVSLEDLEEIVGDIKRCWDERRANMAGGSGRPLPSQSQNATLSTSKPVHSQPPSNDKEPKYVDEDDWSD F7F5A3E59EEFA15F F7F5A3E59EEFA15F cet1_yeast mRNA-capping enzyme subunit beta CET1 mRNA 5'-phosphatase mRNA 5'-triphosphate monophosphatase P1433 YPL228W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSYTDNPPQTKRALSLDDLVNHDENEKVKLQKLSEAANGSRPFAENLESDINQTETGQAAPIDNYKESTGHGSHSQKPKSRKSSNDDEETDTDDEMGASGEINFDSEMDFDYDKQHRNLLSNGSPPMNDGSDANAKLEKPSDDSIHQNSKSDEEQRIPKQGNEGNIASNYITQVPLQKQKQTEKKIAGNAVGSVVKKEEEANAAVDNIFEEKATLQSKKNNIKRDLEVLNEISASSKPSKYRNVPIWAQKWKPTIKALQSINVKDLKIDPSFLNIIPDDDLTKSVQDWVYATIYSIAPELRSFIELEMKFGVIIDAKGPDRVNPPVSSQCVFTELDAHLTPNIDASLFKELSKYIRGISEVTENTGKFSIIESQTRDSVYRVGLSTQRPRFLRMSTDIKTGRVGQFIEKRHVAQLLLYSPKDSYDVKISLNLELPVPDNDPPEKYKSQSPISERTKDRVSYIHNDSCTRIDITKVENHNQNSKSRQSETTHEVELEINTPALLNAFDNITNDSKEYASLIRTFLNNGTIIRRKLSSLSYEIFEGSKKVM 2B946CC53D66F824 2B946CC53D66F824 ceg1-cets-1 2 3 unspecified role unspecified role neutral component neutral component 4 unspecified role unspecified role neutral component neutral component physical association physical association 3 The two bands, at 52 kDa and 75 kDa, were excised from the gel, and the proteins were identified by a combination of peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS) and mass spectrometric sequencing using NanoES triple-quadrupole MS/MS. These analyses unambiguously identified the 52 kDa band as Ceg1p, the guanylyltransferase subunit of the yeast pre-mRNA capping enzyme, and the 75 kDa band as Cet1p, the RNA 5′-triphosphatase subunit of the yeast pre-mRNA capping enzyme. ceg1-cets-2 1 4 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey physical association physical association 5 A crude DE400 fraction was incubated with anti-Cet1p antibodies immobilized on protein-A beads, and the supernatant was recovered. An equal volume of load (DE400) and supernatant (DE400[ΔCet1/Ceg1]) was analyzed by Western blotting.