Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/10giovannicombinationswissprothuman29_06_10.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/13giovanniswissprothuman01_06_10.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/9giovannicombinationswissprothuman29_06_10.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/14giovanniswissprothuman01_07_10.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/10giovannicombinationswissprothuman29_06_10.pdfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/13giovanniswissprothuman01_06_10.pdfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/9giovannicombinationswissprothuman29_06_10.pdfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/14giovanniswissprothuman01_07_10.pdfftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C5.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C2.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C5.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-2.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C3.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-2.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni14-5.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-1.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-4.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C2.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-3.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C6.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C2.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C3.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-3.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-2.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni14-6.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C3.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C4.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-6.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C1.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C6.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-2.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-3.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C5.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C1.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-1.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-4.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-1.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-4.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C3.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C6.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C5.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-4.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C2.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-6.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni14-6.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C4.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-5.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C4.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C6.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-1.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C1.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100702Giovanni14-3.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni13-5.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100630Giovanni14-5.wiffftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100628Giovanni10C1.wiff.scanftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/03/PXD001202/100627Giovanni9C4.wiffprimaryOK200004320Tuula NymanNot availablePermanent Cell Line CellCell CultureHuman mammary carcinoma MCF-7 Tet-Off (Clontech-Takara, Saint-Germain-en-Laye, France) cell lines were used to generate clones stably expressing either C-TAP-ERα, N-TAP-ERα, (indicated globally as TAP-ERα), C-TAP-ERβ or N-TAP-ERβ (indicated as TAP-ERb).TAP tag alone containing clone was used as control. TAP-purification, protein separation by SDS-PAGE and identification by LC-MS/MS was done as previously described. references: Ambrosino, C., Tarallo, R., Bamundo, A., Cuomo, D., et al., Identification of a hormone-regulated dynamic nuclear actin network associated with estrogen receptor alpha in human breast cancer cell nuclei. Mol Cell Proteomics 2010, 9, 1352-1367 Tarallo, R., Bamundo, A., Nassa, G., Nola, E., et al., Identification of proteins associated with ligand-activated estrogen receptor alpha in human breast cancer cell nuclei by tandem affinity purification and nano LC-MS/MS. Proteomics 2011, 11, 172-179. Nassa, G., Tarallo, R., Ambrosino, C., Bamundo, A., et al., A large set of estrogen receptor beta-interacting proteins identified by tandem affinity purification in hormone-responsive human breast cancer cell nuclei. Proteomics 2011, 11, 159-165. Cirillo, F., Nassa, G., Tarallo, R., Stellato, C., et al., Molecular mechanisms of selective estrogen receptor modulator activity in human breast cancer cells: identification of novel nuclear cofactors of antiestrogen-ERalpha complexes by interaction proteomics. J Proteome Res 2013, 12, 421-431.PrideTuula NymanQSTARInstitute of Biotechnology, University of HelsinkiPARTIAL25604459 Stellato C, Nassa G, Tarallo R, Giurato G, Ravo M, Rizzo F, Marchese G, Alexandrova E, Cordella A, Baumann M, Nyman TA, Weisz A, Ambrosino C. Identification of cytoplasmic proteins interacting with unliganded estrogen receptor α and β- in human breast cancer cells. Proteomics. 2015 Jan 20 10.1002/pmic.201400404University of Oslotuula.nyman@helsinki.fiAffinity purification coupled with mass spectrometry proteomicsMass SpectrometryBreast CancerEstrogen receptorTap-tagLc-ms/mshttp://www.ebi.ac.uk/pride/archive/projects/PXD001202Not availablephosphorylated residueiodoacetamide derivatized residuemonohydroxylated residueMS data were acquired using Analyst QS 2.0 software. The LC-MS/MS data were searched against SwissProt 2010 (517802 sequences; 182492287 residues, (human, 20283 sequences) and SwissProt 2010x (516603 sequences; 181919312 residues, human 20278 sequences). The search criteria for Mascot searches were: trypsin digestion with one missed cleavage allowed, Carbamidomethyl (C) as fixed modification and Oxidation (M), Phospho (ST), Phospho (Y) as variable modifications. For the LC-MS/MS spectra the maximum precursor ion mass tolerance was 50 ppm and MS/MS fragment ion mass tolerance 0.2 Da, and peptide charge state of +1, +2 or +3 was used. All of the reported protein identifications were statistically significant (p<0.05). To eliminate the redundancy of proteins that appear in the database under different names and accession numbers, the single protein member with the highest protein score (top rank) was selected from multiprotein families to the identification results.ProteomicsHomo Sapiens (human)tuula.nyman@medisin.uio.noBiomedicalNorwayEstrogen receptor subtypes (ERα and ERβ) are transcription factors sharing a similar structure but exerting opposite roles in breast cancer cells. Besides the well-characterized genomic actions of nuclear ERs upon ligand binding, specific actions of ligand-free ERs in the cytoplasm also affect cellular functions. The identification of cytoplasmic interaction partners of unliganded ERα and ERβ may help characterize the molecular basis of the extra-nuclear mechanism of action of these receptors, revealing novel mechanisms to explain their role in breast cancer response or resistance to endocrine therapy. To this aim, cytoplasmic extracts from human breast cancer MCF-7 cells stably expressing tandem affinity purification-tagged ERα and ERβ and maintained in estrogen-free medium were subject to affinity-purification and MS analysis, leading to the identification of 84 and 142 proteins associated with unliganded ERα and ERβ, respectively. Functional analyses of ER subtype-specific interactomes revealed significant differences in the molecular pathways targeted by each receptor in the cytoplasm. This work, reporting the first identification of the unliganded ERα and ERβ cytoplasmic interactomes in breast cancer cells, provides novel experimental evidence on the nongenomic effects of ERs in the absence of hormonal stimulus. All MS data have been deposited in the ProteomeXchange with identifier PXD001202 (http://proteomecentral.proteomexchange.org/dataset/PXD001202).Identification of cytoplasmic proteins interacting with unliganded estrogen receptor α and β in human breast cancer cells.Stellato Claudia C, Nassa Giovanni G, Tarallo Roberta R, Giurato Giorgio G, Ravo Maria M, Rizzo Francesca F, Marchese Giovanna G, Alexandrova Elena E, Cordella Angela A, Baumann Marc M, Nyman Tuula A TA, Weisz Alessandro A, Ambrosino Concetta CTet, DmelCG5178, FLN-B, SCT, DmelCG7478, Tandem, thyroid autoantigen, Abcb2, Receptor Modulator, SDH, thymoma, xlERalpha1, xlERalpha2, Ham1, DmelCG12051, 5730420M11Rik, dmTAF[[II]]230, SDS, protein polypeptide chains, Ly6, ABP-280 homolog, B1, Line, l(2)SH1330, cancer of the breast, myd, adult, thymic epithelial tumour, A, Nucleus, BC, C, SET, D, E, AU041214, cyt5C, TFIID TAF250, cel, M, Estradiol, phosphatase 2A inhibitor I2PP2A, Mbp-1, proteins, act88F, breast adenocarcinoma cell, endocrine, DmelCG4299, CA - Carcinoma of breast, set, ESR-BETA, Tandem Affinity, cancer of breast, Chromatography, n, Act, act87E, Estradiol Receptor, LC-MS2, dTAF[[II]]230, Ligand, Ly-6A.2, Actin/BAP47, Tandem Affinity Chromatographies, bHLHc9, TAF200, act, act42A, Cell Nuclei, RING4, l(2)br3, epithelial neoplasm of the Thymus, Tap-1, AFFX-Dros-ACTIN_M_r_at, mKIAA0609, ER, Proteomes., actin, Mammary carcinoma, Act57A, Carcinomas, fg, DmelCG7659, DmelCG4027, Antiestrogen, HLA-DR-associated protein II, DI-2, FLN3, mammary cancer cell, I-2Dm, expanded, alpha, inhibitor of granzyme A-activated DNase, FLNB, MEX67, Act57b, human, act79B, I-2PP1, MCF7, MDC1D, actin5C, TAF-IBETA, enr, breast cancer, Taf250, liquid chromatography-tandem mass spectroscopy, Mex67, TAF-Ibeta, cancer, carcinoma of the breast, dipeptide chemoreceptor protein, T-cell-activating protein, l(3)01658, Estradiol Receptor alpha, TAF230, big, F10B6_15, Act5c, Estrogen Receptor, epithelial tumor of Thymus, Peptidomics, Protein USO1 homolog, Tandem Affinity Chromatography, ACT5C, AA420328, Stem cell antigen 1, protein-containing complex, DmelCG4482, F10B6.15, Human, CG30294, large, TAP1, Mex67h, LC-MSMS, Gene Products, Carcinoma of the Breast, Erb, mammary adenocarcinoma cell, ABP-280-like protein, dTAF[[II]]250, cell, MDDGB6, ligand, Mammary Carcinoma, epithelial tumour of Thymus, mus79B, 4733401P19Rik, 2pp2a, ERa, Act57, MCP-IV, man, CG10574, nr3a1, anon-EST:fe2D2, dTAF250, ESR, 2PP2A, CG18290, xesr1, dSET, dSet, T10, T11, Ly-6A.2|Ly-6E.1, truncated ABP, Squalene transfer protein, Act-5C, Ly6a, MCF-7 cell, C22orf6, Receptor alpha, 42A, SPF, Spf, CG4482, Receptor beta, carcinoma of breast, Estrogen Receptors alpha, Proteins, Ly-6E.1, Sca-1, BG:DS00004.13, Cell, dTAF230, Estrogen Receptor 2, Estrogen Receptor 1, Estrogen Receptors beta, native protein, I-2PP2A, beta-actin, Dm I-2, M32055, TAF[[II]]250/230, Human Mammary Carcinoma, Act42, Taf[[II]]250, Estrogen, Thymus epithelial tumor, Alpha-tocopherol-associated protein, Act88-F, PSF1, epithelial neoplasm of Thymus, SWDS, AOI, Supernatant protein factor, mus87E, br3, Gene Proteins, SCA-1, bps, ESTRB, breast tumor cell, MTP1, liquid chromatography tandem mass spectrometry, beta-filamin, BAP47, NIP, Vesicle-docking protein, Bap47, General activity, breast, epithelial tumour of the Thymus, liquid chromatography tandem mass spectroscopy, IPP2A2, act 87E, Affinity Purification, Activity, Mbp1, Ly-6A|E, nip, protein, Estra, ER[a], Estrogen Receptor Modulator, Act5, l(1)G0420, act 88F, hTAP, protein aggregate, xesr-1, rsd, Sca1, LCMSMS, reference sample, TAF-I, Modulators, Human Mammary Carcinomas, breast epithelial cancer cell, breast carcinoma cell, 35Bb, ABC17, thymic epithelium neoplasm, MCF7 cell, IGAAD, LRS1, SOLO DANCERS, DmelCG10574, act 42A, ERbeta, Ac5C, CG7659, CG4027, Carcinoma of breast (disorder), epithelial tumor of the Thymus, stem cell antigen 1, DmelCG18290, associated, filamin-3, 115kDa, Saint, Nr3a1, phapii, Carcinoma, l(1)G0330, gyltl1b-b, Nuclei, LC-MS/MS, AI836084, StF-IT-1, TAFII-250, TAF250/230, ACTIN, TAFII250, l35Bb, MDDGA6, NOS, breast carcinoma, CG7478, CG5178, CG10067, Lines, gyltl1b, AI256582, l(1)G0117, actin-binding-like protein, carcinoma OF breast, mdc1d, APT1, Actin, CG4299, CG17603, TAF[[II]], Saint Pierre and Miquelon, beta-actin/Bap47, l(2)SH2 1330, l(1)G0486, Receptor Modulators, l(1)G0245, l(1)G0009, enlarged, SR3-5, Thymus epithelial tumour, St. Pierre and Miquelon, BG:DS01219.1, Y3, Estrogen receptor alpha, i2pp2a, Affinity Chromatography, act57b, esr1, XER, Carcinoma of breast NOS, d230, l(1)G0010, Act87e, human being, truncated actin-binding protein, Ly-6A.2/Ly-6E.1, mol, Antiestrogens, esr1b, esr1a, CG12051, Gene, dTAFII250, anon-EST:CL2c12, EfW1, 1300013M05Rik, froggy, PHAPII, LC-MS-MS, Gyltl1a, act57B, l(1)G0025, ERbetacx, Modulator, act57A, Act88f, polypeptide chain, template-activating factor I, dmTAF1, Taf230, Miquelon and St. Pierre, actin-12, NR3A2, AW536553, Ham-1, mammary carcinoma cell, 143060_f_at, l(2)br23, TAF250, Taf200, Human Mammary, hormones, act5C, Ly-6A/E, DmelCG10067, Carcinoma of breast NOS (disorder), thymic epithelial tumor, ipp2a2, ERalpha, Taf1p, mammary carcinoma, thymus epithelial neoplasm, TABP, Miquelon and Saint Pierre, Act42a, BPFD#36, Vdp, VDP, taf-ibeta, Thymus epithelial neoplasm, great, mammary tumor cell, TAF, filamin homolog 1, FLN1L, ESRB, Controlled, l(1)G0177, Ifm(3)7, TAP, Tap, ABP-278, ER-BETA, Controlling, TAF[[II]]250, protein complex, act 79B, Fh1, l(2)35Bb, igaad, Cell Lines, total expressed protein, l(3)84Ab, mammary gland carcinoma cell, BEST:RE38067, tap, LC/MS/MS, natural protein, Mammary Carcinomas, p230, FH1, Protein, I2PP2A, TFIID, Mvb1, Nassa <Nassa>, Corsica, ABP-280, l(1)G0079, 10^[-9], TAF[[II]]230, act 57A, act 5C, TAF[II]250, CG15268, CGI-97, P115, Protein Gene Products, dSET/TAF-Ibeta, 2610030F17Rik, DmelCG17603, Transcytosis-associated protein, Act(88F), Estr, responsive, AA407739, TEN, Actin5C, ER-alpha, TAF1BC, human being, carcinoma of breast, NR3A1, nuclear receptor subfamily 3 group A member 1, Proteins, Gene, proteins, mammary carcinoma, Estra, man, human, Protein Gene Products, Gene Proteins, estradiol receptor, ESR, breast cancer, cancer of breast, Estr, ER, ESR1, Protein, Gene Products, NOS, Cell., cancer of the breast, breast carcinoma, carcinoma of the breast, cancer, ER-alpha, breastliquid chromatography tandem mass spectroscopy, PDB2, results., human being, RANK, AW549739, cleavage, FBN, Gene, TRANCER, protein, Computer, precursor, protein-containing complex, LC-MS-MS, peptide, Polypeptides, protein polypeptide chains, Drug Tolerance, ClvPrd, peptido, polypeptide chain, ECTOL1, LC-MSMS, Gene Products, Software Engineering, Isonymies, protein aggregate, Computer Program, WMS, Application, C79691, mRANK, Open Source Softwares, ODFR, OSTS, LCMSMS, peptides, TNFRSF11A, Ly109, Software Application, receptor activator of NF-KB, Open, OFE, beta-Trypsin, Open Source Software, Computer Programs and Programming, proteins, man, Computer Software Application, Lccp, OCTD, mKIAA0989, Tools, 10^[-6], Rank, ppm, FEO, peptidos, OPTB7, GPHYSD2, z, drug tolerance, SGS, Applications Software, LC-MS2, Open Source, data, Computer Software, protein complex, immune system tolerance, Name, Proteins, LC-MS/MS, LOH18CR1, not genetically inherited, Source Softwares, Peptide, Software Tools, Tool, Programs, ACMICD, polypeptide, Program, Computer Applications, Software Tool, LC/MS/MS, TRANCE-R, Immune Tolerance, native protein, Isonymy, natural protein, Computer Applications Software, Protein, Computer Applications Softwares, Immunologic Tolerance, MFS1, Softwares, Software, WMS2, Data Base, CD265, parent ion, Software Applications, Source Software, Engineering, Tripcellim, MASCOT, MASS, beta Trypsin, osteoclast differentiation factor receptor, human, Protein Gene Products, Computer Programs, Trypure, Gene Proteins, Applications, Applications Softwares, Self Tolerance, precursor ion, Tolerance, liquid chromatography-tandem mass spectroscopy, SSKS, liquid chromatography tandem mass spectrometry, Peptid, Polypeptide, variable, Computer Software Applications, Immunological Tolerancemammary region, FLN-B, SCT, Breasts, conformation, HSN1E, truncated actin-binding protein, experimental, Ly-6A.2/Ly-6E.1, thyroid autoantigen, Peptidomics, Protein USO1 homolog, nuclear receptor subfamily 3 group A member 1, Abcb2, estrogenes, Ly-6A|E, EA1, Gene, Estra, Tumor, MLR-3, Stem cell antigen 1, Transcription Factor, 1300013M05Rik, Cell Growth in Number, Ham1, Protoplasm, estradiol receptor, AIM, Ly6, TAP1, Roles, hTAP, ABP-280 homolog, Mex67h, responsivity, resistance, Cell Number Growth, Gene Products, Concepts, AW536553, cancer of the breast, Number Growth, early activation antigen CD69, Ham-1, BC, treatment, reactivity, study, Transcription, Growth, hormones, Ly-6A/E, ABP-280-like protein, methods, Sca1, absent from organism., activation inducer molecule, experimental section, NR3A1, ligand, absent from organism, mammary part of chest, mammary carcinoma, proteins, W, MCP-IV, ABC17, estrogenos, free, TABP, endocrine, MCF7 cell, Vdp, VDP, cell proliferation, ESR, leukocyte surface antigen Leu-23, LRS1, cancer of breast, ESR1, Role Concepts, disease management, oestrogene, C-type lectin domain family 2 member C, CG7659, oestrogens, stem cell antigen 1, Ly-6A.2|Ly-6E.1, associated, filamin-3, Estrogene, truncated ABP, relational structural quality, 115kDa, Squalene transfer protein, filamin homolog 1, Ly6a, FLN1L, C22orf6, MCF-7 cell, MCF 7 cell, mamma, Multiplication, TAP, Tap, ABP-278, Ligand, Cellular Proliferation, Ly-6A.2, SPF, Spf, carcinoma of breast, bHLHc9, Fh1, Cytoplasms, Proteins, DNMT, Oestrogene, Ly-6E.1, Sca-1, estrogene, Factor, MCMT, Cell, Concept, RING4, BL-AC/P26, tap, Role Concept, estrogens, Tap-1, estrogeno, ER, FH1, Protein, Role, Cellular, Oestrogen, NOS, breast carcinoma, Mvb1, ABP-280, CXXC9, oestrogen, early T-cell activation antigen p60, ADCADN, DmelCG7659, absence, Factors, Alpha-tocopherol-associated protein, PSF1, FLN3, estrogenes Hormon, AI256582, actin-binding-like protein, APT1, FLNB, AOI, Supernatant protein factor, MEX67, experimental procedures, P115, Protein Gene Products, Gene Proteins, MCF7, SCA-1, bps, Transcytosis-associated protein, breast cancer, GP32/28, Proliferation, MTP1, Estr, Mex67, Cell Multiplication, Protoplasms, Cell Number, beta-filamin, response, Vesicle-docking protein, carcinoma of the breast, cancer, Y3, CD69, dipeptide chemoreceptor protein, T-cell-activating protein, l(3)01658, CLEC2C, ER-alpha, breast, oestrogenesMCF-7 Cell, Affinity Purification, human being, conformation, HSN1E, Tandem, determination, experimental, AI461847, Tandem Affinity Chromatography, nuclear receptor subfamily 3 group A member 1, MCF7 Cell, estrogenes, EA1, Gene, Estra, MLR-3, Transcription Factor, MCF7 Cells, Protoplasm, estradiol receptor, AIM, Roles, responsivity, resistance, Gene Products, Concepts, Mood, Pgi, cancer of the breast, Gpi-1, early activation antigen CD69, BC, treatment, reactivity, Transcription, hormones, MCF-7, methods, activation inducer molecule, Gpi-1r, Nlk, Moods, experimental section, Gpi-1s, NR3A1, ligand, absent from organism, Phi, Gpi-1t, mammary carcinoma, proteins, Michigan Cancer Foundation 7 Cells, W, estrogenos, man, free, endocrine, ESR, leukocyte surface antigen Leu-23, Tandem Affinity, cancer of breast, ESR1, Role Concepts, Chromatography, disease management, oestrogene, C-type lectin domain family 2 member C, oestrogens, associated, Estrogene, relational structural quality, NK/GPI, Data Set., Gpi, data, Ligand, Tandem Affinity Chromatographies, carcinoma of breast, Cytoplasms, Affects, Proteins, DNMT, Oestrogene, estrogene, Factor, MCF 7 Cells, MCMT, MF, Amf, Cell, Concept, BL-AC/P26, Role Concept, estrogens, mOC-X, estrogeno, ER, chemical analysis, Protein, Role, Oestrogen, NOS, breast carcinoma, NK|GPI, NK, CXXC9, oestrogen, early T-cell activation antigen p60, ADCADN, ORG, Org, absence, Factors, estrogenes Hormon, Gpi1-r, Gpi1-s, Gpi1-t, human, experimental procedures, Protein Gene Products, Gene Proteins, MCF7, breast cancer, GP32/28, Estr, Protoplasms, Cells, assay, response, carcinoma of the breast, cancer, CD69, CLEC2C, Affinity Chromatography, Bglap-rs1, ER-alpha, breast, oestrogenes, Gpi1sBC, human being, carcinoma of breast, NR3A1, nuclear receptor subfamily 3 group A member 1, Proteins, Gene, proteins, mammary carcinoma, Estra, man, human, Protein Gene Products, Gene Proteins, estradiol receptor, ESR, breast cancer, cancer of breast, Estr, ER, ESR1, Protein, Gene Products, NOS, Cell., cancer of the breast, breast carcinoma, carcinoma of the breast, cancer, ER-alpha, breast432trueIdentification of cytoplasmic proteins interacting with unliganded estrogen receptor α and β- in human breast cancer cellsEstrogen Receptor subtypes (ERα and ERβ) are transcription factors sharing similar structure, however, they often perform opposite roles in breast cancer’s cell proliferation and tumor progression. Besides the well-characterized genomic actions of ERs upon ligand binding, rapid non-genomic cytoplasmic changes together with the recently discovered ligand-free action of ERs are emerging as key regulators of tumorigenesis. The identification of cytoplasmic interaction partners of unliganded ERα and ERβ may help characterize the molecular basis of the extra-nuclear mechanism of action of these receptors, revealing novel mechanisms to explain their role in breast cancer response or resistance to endocrine therapy. To this aim, in this study, cytoplasmic extracts from stably expressing TAP-ERα and -ERβ MCF-7 cell clones were subjected to interaction proteomics in the absence of estrogen stimulation, leading to the identification of 84 and 142 proteins associated with unliganded ERα and ERβ, respectively. Functional analyses of ER subtype-specific interactomes revealed significant differences in the molecular pathways associated to each receptor in the cytoplasm. This work reports the first identification of the unliganded ERα and ERβ cytoplasmic interactomes in breast cancer cells, providing novel experimental evidence on the non-genomic effects of ERs in the absence of hormonal 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