bhf-ucl qin-2004-6 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. in vitro In vitro pull down pull down autoradiography autoradiography Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-3 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). gal4 vp16 complement gal4 vp16 complementation karyoplasmic interaction ion strategy KISS mammalian two hybrid gal4 vp16 complement predetermined predetermined participant predetermined Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-1 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins The nine positive clones were verified by retransformation into a yeast strain producing the bait protein before being sequenced. hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Human adult liver cDNA library Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-2 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-4 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-5 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. chlae-cos Chlorocebus aethiops cos cos confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions qin-2004-7 Prospero-related homeobox (Prox1) is a corepressor of human liver receptor homolog-1 and suppresses the transcription of the cholesterol 7-alpha-hydroxylase gene. in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa predetermined predetermined participant predetermined Qin J., Gao D.M., Jiang Q.F., Zhou Q., Kong Y.Y., Wang Y., Xie Y.H. Cardiac - Interactions involving cardiac related proteins hLRH-1 used in this paper is the isoform 2 in UniProt (o00482-2) according to the length of the protein depicted in Fig. 1 yhxie@sibs.ac.cn or wangy@sibs.ac.cn Mol. Endocrinol. (0888-8809) 2004 imex curation Only protein-protein interactions prox1_human Prospero homeobox protein 1 PROX1 Homeobox prospero-like protein PROX1 protein protein human Homo sapiens Human MPDHDSTALLSRQTKRRRVDIGVKRTVGTASAFFAKARATFFSAMNPQGSEQDVEYSVVQHADGEKSNVLRKLLKRANSYEDAMMPFPGATIISQLLKNNMNKNGGTEPSFQASGLSSTGSEVHQEDICSNSSRDSPPECLSPFGRPTMSQFDMDRLCDEHLRAKRARVENIIRGMSHSPSVALRGNENEREMAPQSVSPRESYRENKRKQKLPQQQQQSFQQLVSARKEQKREERRQLKQQLEDMQKQLRQLQEKFYQIYDSTDSENDEDGNLSEDSMRSEILDARAQDSVGRSDNEMCELDPGQFIDRARALIREQEMAENKPKREGNNKERDHGPNSLQPEGKHLAETLKQELNTAMSQVVDTVVKVFSAKPSRQVPQVFPPLQIPQARFAVNGENHNFHTANQRLQCFGDVIIPNPLDTFGNVQMASSTDQTEALPLVVRKNSSDQSASGPAAGGHHQPLHQSPLSATTGFTTSTFRHPFPLPLMAYPFQSPLGAPSGSFSGKDRASPESLDLTRDTTSLRTKMSSHHLSHHPCSPAHPPSTAEGLSLSLIKSECGDLQDMSEISPYSGSAMQEGLSPNHLKKAKLMFFYTRYPSSNMLKTYFSDVKFNRCITSQLIKWFSNFREFYYIQMEKYARQAINDGVTSTEELSITRDCELYRALNMHYNKANDFEVPERFLEVAQITLREFFNAIIAGKDVDPSWKKAIYKVICKLDSEVPEIFKSPNCLQELLHE o00482-2 Nuclear receptor subfamily 5 group A member 2 CPF FTF Alpha-1-fetoprotein transcription factor B1-binding factor CYP7A promoter-binding factor Hepatocytic transcription factor Liver receptor homolog 1 NR5A2 B1F protein protein human Homo sapiens Human MSSNSDTGDLQESLKHGLTPIVSQFKMVNYSYDEDLEELCPVCGDKVSGYHYGLLTCESCKGFFKRTVQNNKRYTCIENQNCQIDKTQRKRCPYCRFQKCLSVGMKLEAVRADRMRGGRNKFGPMYKRDRALKQQKKALIRANGLKLEAMSQVIQAMPSDLTISSAIQNIHSASKGLPLNHAALPPTDYDRSPFVTSPISMTMPPHGSLQGYQTYGHFPSRAIKSEYPDPYTSSPESIMGYSYMDSYQTSSPASIPHLILELLKCEPDEPQVQAKIMAYLQQEQANRSKHEKLSTFGLMCKMADQTLFSIVEWARSSIFFRELKVDDQMKLLQNCWSELLILDHIYRQVVHGKEGSIFLVTGQQVDYSIIASQAGATLNNLMSHAQELVAKLRSLQFDQREFVCLKFLVLFSLDVKNLENFQLVEGVQEQVNAALLDYTMCNYPQQTEKFGQLLLRLPEIRAISMQAEEYLYYKHLNGDVPYNNLLIEMLHAKRA cp7a1_human_gene Cholesterol 7-alpha-monooxygenase CYP7A1 CYP7 CYPVII Cholesterol 7-alpha-hydroxylase Cytochrome P450 7A1 gene gene human Homo sapiens Human nr5a2-prox1-14 1513028 n/a 1513035 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 1513036 unspecified role unspecified role bait bait n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false O00482-2:p.[Gln273Ala;Gln281Ala;Gln282Ala] 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 certain certain sequence position certain certain certain sequence position certain false O00482-2:p.[Lys301Ala;Gln305Ala;Phe308Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O00482-2:p.[Asp326Ala;Asp327Ala;Met329Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O00482-2:p.[Leu453Ala;Leu454Ala] 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 lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 3D nr5a2-prox1-16 1513028 n/a 1513036 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1513035 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro physical association physical association false false false Fig 4A The interaction with GST-hLRH-1 186-495 is dramatically reduced when they use Prox1 fragments 76-333 and 55-253 nr5a2-prox1-17 1513028 n/a 1513036 unspecified role unspecified role bait bait lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false dbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1513035 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 3B nr5a2-prox1-6 1513029 n/a 1513036 unspecified role unspecified role bait bait lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false dbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1513035 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 2C, 3A, 4C nr5a2-prox1-15 1513029 n/a 1513036 unspecified role unspecified role bait bait hinge-lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1513035 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind 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 physical association physical association false false false Fig. 4B nr5a2-prox1-2 1513030 n/a 1513036 unspecified role unspecified role bait bait hinge-lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1513035 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig.1 nr5a2-prox1-4 1513031 n/a 1513036 unspecified role unspecified role bait bait n/a 1513035 unspecified role unspecified role prey prey c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig. 2B nr5a2-prox1-9 1513032 n/a 1513036 unspecified role unspecified role neutral component neutral component n/a 1513035 unspecified role unspecified role neutral component neutral component c-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig. 2D nr5a2-prox1-11 1513033 n/a 1513035 unspecified role unspecified role neutral component neutral component c-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1513036 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig 2D cyp7a1-nr5a2 1513034 n/a 1513037 unspecified role unspecified role bait bait promoter region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false A short fragment of the human cyp7a1 promoter (-122 to -140) was used in the EMSA 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false synthetic construct synthetic construct n/a 1513036 unspecified role unspecified role prey prey rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes physical association physical association false false false Fig. 7 Prox 1 (it binds to LRH blocking the DNA protein interaction)