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/ Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) 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 predetermined predetermined participant predetermined Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! The p53-SV4 positive control interaction has not been added as no origin of p53 is stated. hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) 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 Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS MCF-7 breast cancer cell cDNA library BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. Keratin 18 attenuates estrogen receptor alpha-mediated signaling by sequestering LRP16 in cytoplasm. BMC cell biology 2008 imex curation Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Meng Y., Wu Z., Yin X., Zhao Y., Chen M., Si Y., Yang J., Fu X., Han W. Cancer - Interactions investigated in the context of cancer "pcDNA3-Flag" - Tag appears to be N-terminal based on looking at ~10 of 42 of these plasmids from Addgene! hanwdrsw69@yahoo.com Accepted 2018-JUN-25 AT 10:04 BST AT 10:04 BST by PPORRAS BMC cell biology 2008 Only protein-protein interactions imex curation imex curation done k1c18_human Keratin, type I cytoskeletal 18 KRT18 CYK18 Keratin-18 PIG46 Cytokeratin-18 Cell proliferation-inducing gene 46 protein protein protein human Homo sapiens Human MSFTTRSTFSTNYRSLGSVQAPSYGARPVSSAASVYAGAGGSGSRISVSRSTSFRGGMGSGGLATGIAGGLAGMGGIQNEKETMQSLNDRLASYLDRVRSLETENRRLESKIREHLEKKGPQVRDWSHYFKIIEDLRAQIFANTVDNARIVLQIDNARLAADDFRVKYETELAMRQSVENDIHGLRKVIDDTNITRLQLETEIEALKEELLFMKKNHEEEVKGLQAQIASSGLTVEVDAPKSQDLAKIMADIRAQYDELARKNREELDKYWSQQIEESTTVVTTQSAEVGAAETTLTELRRTVQSLEIDLDSMRNLKASLENSLREVEARYALQMEQLNGILLHLESELAQTRAEGQRQAQEYEALLNIKVKLEAEIATYRRLLEDGEDFNLGDALDSSNSMQTIQKTTTRRIVDGKVVSETNDTKVLRH DIP protein P05783 original sequence version: 186 DIP protein P05783 original sequence version: 175 1E5604C6BCC7A17A tff1_human_gene PS2/TFFI HUMAN GENE gene gene human Homo sapiens Human esr1_human Estrogen receptor ESR1 ESR NR3A1 Estradiol receptor ER-alpha Nuclear receptor subfamily 3 group A member 1 protein protein human Homo sapiens Human MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV DIP protein P03372 original sequence version: 214 DIP protein P03372 original sequence version: 139 DIP protein P03372 original sequence version: 167 5455C57AB0CCCAA7 macd1_human ADP-ribose glycohydrolase MACROD1 MACROD1 LRP16 Protein LRP16 MACRO domain-containing protein 1 O-acetyl-ADP-ribose deacetylase MACROD1 [Protein ADP-ribosylaspartate] hydrolase MACROD1 [Protein ADP-ribosylglutamate] hydrolase MACROD1 protein protein human Homo sapiens Human MSLQSRLSGRLAQLRAAGQLLVPPRPRPGHLAGATRTRSSTCGPPAFLGVFGRRARTSAGVGAWGAAAVGRTAGVRTWAPLAMAAKVDLSTSTDWKEAKSFLKGLSDKQREEHYFCKDFVRLKKIPTWKEMAKGVAVKVEEPRYKKDKQLNEKISLLRSDITKLEVDAIVNAANSSLLGGGGVDGCIHRAAGPLLTDECRTLQSCKTGKAKITGGYRLPAKYVIHTVGPIAYGEPSASQAAELRSCYLSSLDLLLEHRLRSVAFPCISTGVFGYPCEAAAEIVLATLREWLEQHKDKVDRLIICVFLEKDEDIYRSRLPHYFPVA 82294BFC904FA4D0 macrod1-krt18-1 4 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association no figure macrod1-krt18-2 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Figure 1 macrod1-krt18-3 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed partially purified partially purified region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c2_tail_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed partially purified partially purified 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 2 macrod1-krt18-4 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed partially purified partially purified 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 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed partially purified partially purified 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 macro_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Figure 3 124345qwerry-1 1 7 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed association association Figure 4A krt18-1 6 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range c1_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 4B krt18-macrod1-1 1 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Figure 7C-1 E2 stimulation enhanced the interaction between LRP16 and ERalpha but had no effect on the interaction between K18 and LRP16. krt18-macrod1-2 1 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 7C-2 Ectopic K18 expression in MCF-7 cells markedly attenuated the association of ERalpha with LRP16. ectopic K18 expression in MCF-7 cells markedly attenuated the association of ERα with LRP16 macrod1-esr1-2 1 9 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 7C-3-RHS Cells were cultured in phenol-red free media stripped of steroids for at least 3 days, then treated with E2. E2 stimulation enhanced the interaction between LRP16 and ERalpha but had no effect on the interaction between K18 and LRP16. E2 - Estradiol E2 stimulation enhanced the interaction between LRP16 and ERalpha esr1-tff1_human_gene-1 2 9 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level promoter_region (-353 to -30) sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Figure 8C E2 - Estradiol ERalpha binding at the pS2 promoter was significantly increased in the presence of E2, but binding was substantially blunted by overexpression of K18. ERalpha binding at the pS2 promoter was significantly increased in the presence of E2, but binding was substantially blunted by overexpression of K18. K18 - P05783 CHEBI:23965 increases binding of P03372 to ENSG00000160182 P05783 decreases binding of P03372 to ENSG00000160182 CHEBI:23965 increases expression at RNA level of ENSG00000160182. P05783 decreases expression at RNA level of ENSG00000160182. RNA causality statements from Fig 8 A/B Q9BQ69 reduced the P05783 induced decrease in expression at RNA level of ENSG00000160182.