bhf-ucl song-2006f-2 A Prospero-related homeodomain protein is a novel co-regulator of hepatocyte nuclear factor 4alpha that regulates the cholesterol 7alpha-hydroxylase gene. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast lexa b52 complement lexa b52 complementation LexA B52 transcription complementation lexa b52 complement predetermined predetermined participant predetermined Song K.H., Li T., Chiang J.Y. Cardiac - Interactions involving cardiac related proteins The author has confirmed that Prox1, SHP, SF-1, HNF4-alpha, FTF and PGC1-alpha were all of human origin. jchiang@neoucom.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation song-2006f-4 A Prospero-related homeodomain protein is a novel co-regulator of hepatocyte nuclear factor 4alpha that regulates the cholesterol 7alpha-hydroxylase gene. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Song K.H., Li T., Chiang J.Y. Cardiac - Interactions involving cardiac related proteins The author has confirmed that Prox1, SHP, SF-1, HNF4-alpha, FTF and PGC1-alpha were all of human origin. jchiang@neoucom.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation song-2006f-3 A Prospero-related homeodomain protein is a novel co-regulator of hepatocyte nuclear factor 4alpha that regulates the cholesterol 7alpha-hydroxylase gene. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Song K.H., Li T., Chiang J.Y. Cardiac - Interactions involving cardiac related proteins The author has confirmed that Prox1, SHP, SF-1, HNF4-alpha, FTF and PGC1-alpha were all of human origin. jchiang@neoucom.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation song-2006f-1 A Prospero-related homeodomain protein is a novel co-regulator of hepatocyte nuclear factor 4alpha that regulates the cholesterol 7alpha-hydroxylase gene. human-hepatocytes Human-hepatocytes primary hepatocytes primary hepatocytes anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Song K.H., Li T., Chiang J.Y. Cardiac - Interactions involving cardiac related proteins The author has confirmed that Prox1, SHP, SF-1, HNF4-alpha, FTF and PGC1-alpha were all of human origin. jchiang@neoucom.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation prox1_human Prospero homeobox protein 1 PROX1 Homeobox prospero-like protein PROX1 protein protein human Homo sapiens Human MPDHDSTALLSRQTKRRRVDIGVKRTVGTASAFFAKARATFFSAMNPQGSEQDVEYSVVQHADGEKSNVLRKLLKRANSYEDAMMPFPGATIISQLLKNNMNKNGGTEPSFQASGLSSTGSEVHQEDICSNSSRDSPPECLSPFGRPTMSQFDMDRLCDEHLRAKRARVENIIRGMSHSPSVALRGNENEREMAPQSVSPRESYRENKRKQKLPQQQQQSFQQLVSARKEQKREERRQLKQQLEDMQKQLRQLQEKFYQIYDSTDSENDEDGNLSEDSMRSEILDARAQDSVGRSDNEMCELDPGQFIDRARALIREQEMAENKPKREGNNKERDHGPNSLQPEGKHLAETLKQELNTAMSQVVDTVVKVFSAKPSRQVPQVFPPLQIPQARFAVNGENHNFHTANQRLQCFGDVIIPNPLDTFGNVQMASSTDQTEALPLVVRKNSSDQSASGPAAGGHHQPLHQSPLSATTGFTTSTFRHPFPLPLMAYPFQSPLGAPSGSFSGKDRASPESLDLTRDTTSLRTKMSSHHLSHHPCSPAHPPSTAEGLSLSLIKSECGDLQDMSEISPYSGSAMQEGLSPNHLKKAKLMFFYTRYPSSNMLKTYFSDVKFNRCITSQLIKWFSNFREFYYIQMEKYARQAINDGVTSTEELSITRDCELYRALNMHYNKANDFEVPERFLEVAQITLREFFNAIIAGKDVDPSWKKAIYKVICKLDSEVPEIFKSPNCLQELLHE stf1_human Steroidogenic factor 1 Nuclear receptor subfamily 5 group A member 1 Adrenal 4-binding protein Steroid hormone receptor Ad4BP Fushi tarazu factor homolog 1 NR5A1 AD4BP FTZF1 SF1 protein protein human Homo sapiens Human MDYSYDEDLDELCPVCGDKVSGYHYGLLTCESCKGFFKRTVQNNKHYTCTESQSCKIDKTQRKRCPFCRFQKCLTVGMRLEAVRADRMRGGRNKFGPMYKRDRALKQQKKAQIRANGFKLETGPPMGVPPPPPPAPDYVLPPSLHGPEPKGLAAGPPAGPLGDFGAPALPMAVPGAHGPLAGYLYPAFPGRAIKSEYPEPYASPPQPGLPYGYPEPFSGGPNVPELILQLLQLEPDEDQVRARILGCLQEPTKSRPDQPAAFGLLCRMADQTFISIVDWARRCMVFKELEVADQMTLLQNCWSELLVFDHIYRQVQHGKEGSILLVTGQEVELTTVATQAGSLLHSLVLRAQELVLQLLALQLDRQEFVCLKFIILFSLDLKFLNNHILVKDAQEKANAALLDYTLCHYPHCGDKFQQLLLCLVEVRALSMQAKEYLYHKHLGNEMPRNNLLIEMLQAKQT hnf4a_human Hepatocyte nuclear factor 4-alpha Transcription factor HNF-4 Nuclear receptor subfamily 2 group A member 1 Transcription factor 14 HNF4A HNF4 NR2A1 TCF14 protein protein human Homo sapiens Human MRLSKTLVDMDMADYSAALDPAYTTLEFENVQVLTMGNDTSPSEGTNLNAPNSLGVSALCAICGDRATGKHYGASSCDGCKGFFRRSVRKNHMYSCRFSRQCVVDKDKRNQCRYCRLKKCFRAGMKKEAVQNERDRISTRRSSYEDSSLPSINALLQAEVLSRQITSPVSGINGDIRAKKIASIADVCESMKEQLLVLVEWAKYIPAFCELPLDDQVALLRAHAGEHLLLGATKRSMVFKDVLLLGNDYIVPRHCPELAEMSRVSIRILDELVLPFQELQIDDNEYAYLKAIIFFDPDAKGLSDPGKIKRLRSQVQVSLEDYINDRQYDSRGRFGELLLLLPTLQSITWQMIEQIQFIKLFGMAKIDNLLQEMLLGGSPSDAPHAHHPLHPHLMQEHMGTNVIVANTMPTHLSNGQMCEWPRPRGQAATPETPQPSPPGGSGSEPYKLLPGAVATIVKPLSAIPQPTITKQEVI nr0b2_human Nuclear receptor subfamily 0 group B member 2 NR0B2 SHP Orphan nuclear receptor SHP Small heterodimer partner protein protein human Homo sapiens Human MSTSQPGACPCQGAASRPAILYALLSSSLKAVPRPRSRCLCRQHRPVQLCAPHRTCREALDVLAKTVAFLRNLPSFWQLPPQDQRRLLQGCWGPLFLLGLAQDAVTFEVAEAPVPSILKKILLEEPSSSGGSGQLPDRPQPSLAAVQWLQCCLESFWSLELSPKEYACLKGTILFNPDVPGLQAASHIGHLQQEAHWVLCEVLEPWCPAAQGRLTRVLLTASTLKSIPTSLLGDLFFRPIIGDVDIAGLLGDMLLLR cp7a1_human_gene Cholesterol 7-alpha-monooxygenase CYP7A1 CYP7 CYPVII Cholesterol 7-alpha-hydroxylase Cytochrome P450 7A1 gene gene human Homo sapiens Human prgc1_human Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Ligand effect modulator 6 PPARGC1A LEM6 PGC1 PGC1A PPARGC1 protein protein human Homo sapiens Human MAWDMCNQDSESVWSDIECAALVGEDQPLCPDLPELDLSELDVNDLDTDSFLGGLKWCSDQSEIISNQYNNEPSNIFEKIDEENEANLLAVLTETLDSLPVDEDGLPSFDALTDGDVTTDNEASPSSMPDGTPPPQEAEEPSLLKKLLLAPANTQLSYNECSGLSTQNHANHNHRIRTNPAIVKTENSWSNKAKSICQQQKPQRRPCSELLKYLTTNDDPPHTKPTENRNSSRDKCTSKKKSHTQSQSQHLQAKPTTLSLPLTPESPNDPKGSPFENKTIERTLSVELSGTAGLTPPTTPPHKANQDNPFRASPKLKSSCKTVVPPPSKKPRYSESSGTQGNNSTKKGPEQSELYAQLSKSSVLTGGHEERKTKRPSLRLFGDHDYCQSINSKTEILINISQELQDSRQLENKDVSSDWQGQICSSTDSDQCYLRETLEASKQVSPCSTRKQLQDQEIRAELNKHFGHPSQAVFDDEADKTGELRDSDFSNEQFSKLPMFINSGLAMDGLFDDSEDESDKLSYPWDGTQSYSLFNVSPSCSSFNSPCRDSVSPPKSLFSQRPQRMRSRSRSFSRHRSCSRSPYSRSRSRSPGSRSSSRSCYYYESSHYRHRTHRNSPLYVRSRSRSPYSRRPRYDSYEEYQHERLKREEYRREYEKRESERAKQRERQRQKAIEERRVIYVGKIRPDTTRTELRDRFEVFGEIEECTVNLRDDGDSYGFITYRYTCDAFAALENGYTLRRSNETDFELYFCGRKQFFKSNYADLDSNSDDFDPASTKSKYDSLDFDSLLKEAQRSLRR nr5a2_human Nuclear receptor subfamily 5 group A member 2 Alpha-1-fetoprotein transcription factor Hepatocytic transcription factor B1-binding factor CYP7A promoter-binding factor Liver receptor homolog 1 NR5A2 B1F CPF FTF protein protein human Homo sapiens Human MSSNSDTGDLQESLKHGLTPIGAGLPDRHGSPIPARGRLVMLPKVETEALGLARSHGEQGQMPENMQVSQFKMVNYSYDEDLEELCPVCGDKVSGYHYGLLTCESCKGFFKRTVQNNKRYTCIENQNCQIDKTQRKRCPYCRFQKCLSVGMKLEAVRADRMRGGRNKFGPMYKRDRALKQQKKALIRANGLKLEAMSQVIQAMPSDLTISSAIQNIHSASKGLPLNHAALPPTDYDRSPFVTSPISMTMPPHGSLQGYQTYGHFPSRAIKSEYPDPYTSSPESIMGYSYMDSYQTSSPASIPHLILELLKCEPDEPQVQAKIMAYLQQEQANRSKHEKLSTFGLMCKMADQTLFSIVEWARSSIFFRELKVDDQMKLLQNCWSELLILDHIYRQVVHGKEGSIFLVTGQQVDYSIIASQAGATLNNLMSHAQELVAKLRSLQFDQREFVCLKFLVLFSLDVKNLENFQLVEGVQEQVNAALLDYTMCNYPQQTEKFGQLLLRLPEIRAISMQAEEYLYYKHLNGDVPYNNLLIEMLHAKRA prox1-nr5a1 1537663 n/a 1537667 unspecified role unspecified role bait bait n/a 1537668 unspecified role unspecified role prey prey physical association physical association false false false Table 1 prox1-hnf4a 1537663 n/a 1537667 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false Q92786:p.[Leu70Ala;Leu73Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1537669 unspecified role unspecified role prey prey af2 domain required to bind necessary binding region required to bind deletion analysis deletion analysis range range range range false physical association physical association false false false Table 1, Fig. 2 nr0b2-hnf4a 1537663 n/a 1537669 unspecified role unspecified role prey prey n/a 1537670 unspecified role unspecified role bait bait physical association physical association false false false Table 1 hnf4a-cyp7a1-3 1537664 n/a 1537671 unspecified role unspecified role prey prey bare-l bare-ll promoter region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1537669 unspecified role unspecified role bait bait association association false false false Fig. 5A Flag-Prox1 was transfected into the HepG2 cells but did not affect HNF4 alpha binding to the CYP7A1 promoter. prox1-cyp7a1-2 1537664 n/a 1537671 unspecified role unspecified role prey prey bare-l bare-ll promoter region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1537667 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 false association association false false false Fig. 5A A positive CHIP result was obtained using an anti-FLAG antibody. The authors state:- "Because Prox1 does not bind to DNA, Prox1 must be recruited to CYP7A1 chromatin by HNF4α". However even if true, this does not exclude the possibility that some unknown molecule could be responsible for binding of Flag-Prox1 to the CYP7A1 promoter region. ppargc1a-cyp7a1-2 1537664 n/a 1537671 unspecified role unspecified role prey prey bare-l bare-ll promoter region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1537672 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig. 5B Q92786 decreases activity of Q9UBK2 Flag-Prox1 Q92786 inhibits expression at RNA level of ENSG00000101076 P41235 increases expression at RNA level of ENSG00000167910 Q9UBK2 increases expression at RNA level of ENSG00000167910 prox1-hnf4a-1 1537665 n/a 1537667 unspecified role unspecified role bait bait purified purified 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 false ecoli Escherichia coli (strain K12) n/a 1537669 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 false in vitro In vitro physical association physical association false false false Fig 1A prox1-nr5a2 1537665 n/a 1537673 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 false in vitro In vitro n/a 1537667 unspecified role unspecified role bait bait purified purified 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1A hnf4a-prox1-1 1537666 n/a 1537667 unspecified role unspecified role prey prey n/a 1537669 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1B