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/ Structure and function of the human transcription elongation factor DSIF. Structure and function of the human transcription elongation factor DSIF. J. Biol. Chem. (0021-9258) 1999 imex curation Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp 27-Aug-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip autoradiography autoradiography Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp Accepted 2008-AUG-9 by ORCHARD. 27-Aug-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1999 orchard Only protein-protein interactions imex curation Structure and function of the human transcription elongation factor DSIF. Structure and function of the human transcription elongation factor DSIF. J. Biol. Chem. (0021-9258) 1999 imex curation Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp 27-Aug-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting autoradiography autoradiography Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp Accepted 2008-AUG-9 by ORCHARD. 27-Aug-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1999 orchard Only protein-protein interactions imex curation Structure and function of the human transcription elongation factor DSIF. Structure and function of the human transcription elongation factor DSIF. J. Biol. Chem. (0021-9258) 1999 imex curation Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp 27-Aug-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down autoradiography autoradiography Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp Accepted 2008-AUG-9 by ORCHARD. 27-Aug-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 1999 orchard Only protein-protein interactions imex curation Structure and function of the human transcription elongation factor DSIF. Structure and function of the human transcription elongation factor DSIF. J. Biol. Chem. (0021-9258) 1999 imex curation Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp 27-Aug-2008: Contacted by JYOTI. orchard Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Yamaguchi Y., Wada T., Watanabe D., Takagi T., Hasegawa J., Handa H. Chromatin - Epigenetic interactions resulting in chromatin modulation hhanda@bio.titech.ac.jp Accepted 2008-AUG-9 by ORCHARD. J. Biol. Chem. (0021-9258) 1999 orchard Only protein-protein interactions imex curation spt5h_human Transcription elongation factor SPT5 DRB sensitivity-inducing factor large subunit DRB sensitivity-inducing factor 160 kDa subunit Tat-cotransactivator 1 protein SUPT5H SPT5 SPT5H protein protein human Homo sapiens Human MSDSEDSNFSEEEDSERSSDGEEAEVDEERRSAAGSEKEEEPEDEEEEEEEEEYDEEEEEEDDDRPPKKPRHGGFILDEADVDDEYEDEDQWEDGAEDILEKEEIEASNIDNVVLDEDRSGARRLQNLWRDQREEELGEYYMKKYAKSSVGETVYGGSDELSDDITQQQLLPGVKDPNLWTVKCKIGEERATAISLMRKFIAYQFTDTPLQIKSVVAPEHVKGYIYVEAYKQTHVKQAIEGVGNLRLGYWNQQMVPIKEMTDVLKVVKEVANLKPKSWVRLKRGIYKDDIAQVDYVEPSQNTISLKMIPRIDYDRIKARMSLKDWFAKRKKFKRPPQRLFDAEKIRSLGGDVASDGDFLIFEGNRYSRKGFLFKSFAMSAVITEGVKPTLSELEKFEDQPEGIDLEVVTESTGKEREHNFQPGDNVEVCEGELINLQGKILSVDGNKITIMPKHEDLKDMLEFPAQELRKYFKMGDHVKVIAGRFEGDTGLIVRVEENFVILFSDLTMHELKVLPRDLQLCSETASGVDVGGQHEWGELVQLDPQTVGVIVRLERETFQVLNMYGKVVTVRHQAVTRKKDNRFAVALDSEQNNIHVKDIVKVIDGPHSGREGEIRHLFRSFAFLHCKKLVENGGMFVCKTRHLVLAGGSKPRDVTNFTVGGFAPMSPRISSPMHPSAGGQRGGFGSPGGGSGGMSRGRGRRDNELIGQTVRISQGPYKGYIGVVKDATESTARVELHSTCQTISVDRQRLTTVGSRRPGGMTSTYGRTPMYGSQTPMYGSGSRTPMYGSQTPLQDGSRTPHYGSQTPLHDGSRTPAQSGAWDPNNPNTPSRAEEEYEYAFDDEPTPSPQAYGGTPNPQTPGYPDPSSPQVNPQYNPQTPGTPAMYNTDQFSPYAAPSPQGSYQPSPSPQSYHQVAPSPAGYQNTHSPASYHPTPSPMAYQASPSPSPVGYSPMTPGAPSPGGYNPHTPGSGIEQNSSDWVTTDIQVKVRDTYLDTQVVGQTGVIRSVTGGMCSVYLKDSEKVVSISSEHLEPITPTKNNKVKVILGEDREATGVLLSIDGEDGIVRMDLDEQLKILNLRFLGKLLEA DIP protein O00267 original sequence version: 130 DIP protein O00267 original sequence version: 91 DIP protein O00267 original sequence version: 131 DIP protein O00267-1 original sequence version: 150 DIP protein O00267 original sequence version: 44 EC3F402A670A5B7D rpb1_human DNA-directed RNA polymerase II subunit RPB1 POLR2A DNA-directed RNA polymerase II subunit A DNA-directed RNA polymerase III largest subunit POLR2 RNA-directed RNA polymerase II subunit RPB1 protein protein human Homo sapiens Human MHGGGPPSGDSACPLRTIKRVQFGVLSPDELKRMSVTEGGIKYPETTEGGRPKLGGLMDPRQGVIERTGRCQTCAGNMTECPGHFGHIELAKPVFHVGFLVKTMKVLRCVCFFCSKLLVDSNNPKIKDILAKSKGQPKKRLTHVYDLCKGKNICEGGEEMDNKFGVEQPEGDEDLTKEKGHGGCGRYQPRIRRSGLELYAEWKHVNEDSQEKKILLSPERVHEIFKRISDEECFVLGMEPRYARPEWMIVTVLPVPPLSVRPAVVMQGSARNQDDLTHKLADIVKINNQLRRNEQNGAAAHVIAEDVKLLQFHVATMVDNELPGLPRAMQKSGRPLKSLKQRLKGKEGRVRGNLMGKRVDFSARTVITPDPNLSIDQVGVPRSIAANMTFAEIVTPFNIDRLQELVRRGNSQYPGAKYIIRDNGDRIDLRFHPKPSDLHLQTGYKVERHMCDGDIVIFNRQPTLHKMSMMGHRVRILPWSTFRLNLSVTTPYNADFDGDEMNLHLPQSLETRAEIQELAMVPRMIVTPQSNRPVMGIVQDTLTAVRKFTKRDVFLERGEVMNLLMFLSTWDGKVPQPAILKPRPLWTGKQIFSLIIPGHINCIRTHSTHPDDEDSGPYKHISPGDTKVVVENGELIMGILCKKSLGTSAGSLVHISYLEMGHDITRLFYSNIQTVINNWLLIEGHTIGIGDSIADSKTYQDIQNTIKKAKQDVIEVIEKAHNNELEPTPGNTLRQTFENQVNRILNDARDKTGSSAQKSLSEYNNFKSMVVSGAKGSKINISQVIAVVGQQNVEGKRIPFGFKHRTLPHFIKDDYGPESRGFVENSYLAGLTPTEFFFHAMGGREGLIDTAVKTAETGYIQRRLIKSMESVMVKYDATVRNSINQVVQLRYGEDGLAGESVEFQNLATLKPSNKAFEKKFRFDYTNERALRRTLQEDLVKDVLSNAHIQNELEREFERMREDREVLRVIFPTGDSKVVLPCNLLRMIWNAQKIFHINPRLPSDLHPIKVVEGVKELSKKLVIVNGDDPLSRQAQENATLLFNIHLRSTLCSRRMAEEFRLSGEAFDWLLGEIESKFNQAIAHPGEMVGALAAQSLGEPATQMTLNTFHYAGVSAKNVTLGVPRLKELINISKKPKTPSLTVFLLGQSARDAERAKDILCRLEHTTLRKVTANTAIYYDPNPQSTVVAEDQEWVNVYYEMPDFDVARISPWLLRVELDRKHMTDRKLTMEQIAEKINAGFGDDLNCIFNDDNAEKLVLRIRIMNSDENKMQEEEEVVDKMDDDVFLRCIESNMLTDMTLQGIEQISKVYMHLPQTDNKKKIIITEDGEFKALQEWILETDGVSLMRVLSEKDVDPVRTTSNDIVEIFTVLGIEAVRKALERELYHVISFDGSYVNYRHLALLCDTMTCRGHLMAITRHGVNRQDTGPLMKCSFEETVDVLMEAAAHGESDPMKGVSENIMLGQLAPAGTGCFDLLLDAEKCKYGMEIPTNIPGLGAAGPTGMFFGSAPSPMGGISPAMTPWNQGATPAYGAWSPSVGSGMTPGAAGFSPSAASDASGFSPGYSPAWSPTPGSPGSPGPSSPYIPSPGGAMSPSYSPTSPAYEPRSPGGYTPQSPSYSPTSPSYSPTSPSYSPTSPNYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPSYSPTSPNYSPTSPNYTPTSPSYSPTSPSYSPTSPNYTPTSPNYSPTSPSYSPTSPSYSPTSPSYSPSSPRYTPQSPTYTPSSPSYSPSSPSYSPASPKYTPTSPSYSPSSPEYTPTSPKYSPTSPKYSPTSPKYSPTSPTYSPTTPKYSPTSPTYSPTSPVYTPTSPKYSPTSPTYSPTSPKYSPTSPTYSPTSPKGSTYSPTSPGYSPTSPTYSLTSPAISPDDSDEEN DIP protein P24928 original sequence version: 108 DIP protein P24928 original sequence version: 147 DIP protein P24928 original sequence version: 158 DIP protein P24928 original sequence version: 157 DIP protein P24928 original sequence version: 179 DIP protein P24928 original sequence version: 181 DIP protein P24928 original sequence version: 161 DIP protein P24928 original sequence version: 175 DIP protein P24928-1 original sequence version: 175 DIP protein P24928 original sequence version: 68 DIP protein P24928 original sequence version: 116 28D6FD25693A6472 spt4h_human Transcription elongation factor SPT4 DRB sensitivity-inducing factor small subunit DRB sensitivity-inducing factor 14 kDa subunit SUPT4H1 SPT4H SUPT4H protein protein human Homo sapiens Human MALETVPKDLRHLRACLLCSLVKTIDQFEYDGCDNCDAYLQMKGNREMVYDCTSSSFDGIIAMMSPEDSWVSKWQRVSNFKPGVYAVSVTGRLPQGIVRELKSRGVAYKSRDTAIKT DIP protein P63272 original sequence version: 96 DIP protein P63272 original sequence version: 114 09EDF007501D0F03 p160-p14-1 4 5 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3A HA-p14 was co-precipitated with Flag-p160 (lane 8), demonstrating their interaction in vivo. p14-p160 3 7 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction 3B, 3C, 3D The interaction was unaffected by the presence of 0.5% Nonidet P-40 and 1 M KCl (lane 6). Because p14 contains a putative zinc finger motif conserved among different species (13), we examined the effect of divalent cations (lane 4) and a high concentration of a chelating agent (lane 5). Even under these conditions, p160 efficiently bound to p14, suggesting that the zinc finger is not required for the interaction. p14-p160-1 2 5 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 7 unspecified role unspecified role prey prey region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction 5A Pol II purified from HeLa cell nuclear pellet was separated by SDS-PAGE, transferred to polyvinylidene difluoride membrane, renatured, and probed with 35S-labeled p160. As shown in Fig. 5A, a clear band of ~200 kDa, which co-migrates with the largest subunit of pol II, was detected. p160 also reacted with GST-p14 protein which was similarly blotted onto the membrane. pol_iia-p160-2 2 6 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 5 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 5A Pol II purified from HeLa cell nuclear pellet was separated by SDS-PAGE, transferred to polyvinylidene difluoride membrane, renatured, and probed with 35S-labeled p160. As shown in Fig. 5A, a clear band of ~200 kDa, which co-migrates with the largest subunit of pol II, was detected. p160 also reacted with GST-p14 protein which was similarly blotted onto the membrane. Purified POL IIa used in the experiment. pol_iia-p160-3 1 6 unspecified role unspecified role bait bait human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association 5A Purified POL IIa used in the experiment. Fine mapping, however, revealed that aa 313-420 of p160, an N-terminal region in close proximity to the KOW motifs, is sufficient for pol II binding (lane 23). aa 420-757 of p160, which contains all four of the KOW motifs, also bound to pol II weakly Region from AA 420 - 757 bound weakly to Pol II.