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/ http://www.ebi.ac.uk/ li-2012a-2 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-3 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-4 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-1 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. pull down pull down sequence tag sequence tag identification sequence tag Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-6 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-7 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation li-2012a-8 The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The authors confirm thqt this experiment was carried out in SKOV-3 cells (as stated in the text) - rather than 293t cells (as stated in the image legend). The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation h2ay_human Core histone macro-H2A.1 MACROH2A1 H2AFY Histone H2A.y Medulloblastoma antigen MU-MB-50.205 protein protein human Homo sapiens Human MSSRGGKKKSTKTSRSAKAGVIFPVGRMLRYIKKGHPKYRIGVGAPVYMAAVLEYLTAEILELAGNAARDNKKGRVTPRHILLAVANDEELNQLLKGVTIASGGVLPNIHPELLAKKRGSKGKLEAIITPPPAKKAKSPSQKKPVSKKAGGKKGARKSKKKQGEVSKAASADSTTEGTPADGFTVLSTKSLFLGQKLNLIHSEISNLAGFEVEAIINPTNADIDLKDDLGNTLEKKGGKEFVEAVLELRKKNGPLEVAGAAVSAGHGLPAKFVIHCNSPVWGADKCEELLEKTVKNCLALADDKKLKSIAFPSIGSGRNGFPKQTAAQLILKAISSYFVSTMSSSIKTVYFVLFDSESIGIYVQEMAKLDAN erbb2_human Receptor tyrosine-protein kinase erbB-2 p185erbB2 MLN19 Proto-oncogene c-ErbB-2 Tyrosine kinase-type cell surface receptor HER2 Proto-oncogene Neu ERBB2 HER2 NEU NGL Metastatic lymph node gene 19 protein protein protein human Homo sapiens Human MELAALCRWGLLLALLPPGAASTQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTSIISAVVGILLVVVLGVVFGILIKRRQQKIRKYTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLGPASPLDSTFYRSLLEDDDMGDLVDAEEYLVPQQGFFCPDPAPGAGGMVHHRHRSSSTRSGGGDLTLGLEPSEEEAPRSPLAPSEGAGSDVFDGDLGMGAAKGLQSLPTHDPSPLQRYSEDPTVPLPSETDGYVAPLTCSPQPEYVNQPDVRPQPPSPREGPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLTPQGGAAPQPHPPPAFSPAFDNLYYWDQDPPERGAPPSTFKGTPTAENPEYLGLDVPV o75367-3 Core histone macro-H2A.1 MACROH2A1 H2AFY Histone H2A.y Medulloblastoma antigen MU-MB-50.205 protein protein human Homo sapiens Human MSSRGGKKKSTKTSRSAKAGVIFPVGRMLRYIKKGHPKYRIGVGAPVYMAAVLEYLTAEILELAGNAARDNKKGRVTPRHILLAVANDEELNQLLKGVTIASGGVLPNIHPELLAKKRGSKGKLEAIITPPPAKKAKSPSQKKPVSKKAGGKKGARKSKKQGEVSKAASADSTTEGTPADGFTVLSTKSLFLGQKLNLIHSEISNLAGFEVEAIINPTNADIDLKDDLGNTLEKKGGKEFVEAVLELRKKNGPLEVAGAAVSAGHGLPAKFVIHCNSPVWGADKCEELLEKTVKNCLALADDKKLKSIAFPSIGSGRNGFPKQTAAQLILKAISSYFVSTMSSSIKTVYFVLFDSESIGIYVQEMAKLDAN erbb2_human_gene-1 ERBB2_human_gene gene gene human Homo sapiens Human erbb2-h2afy-1 1307574 n/a 1307581 unspecified role unspecified role prey prey n/a 1307582 unspecified role unspecified role bait bait physical association physical association false false false Fig 1C/D h2afy-erbb2-2 1307574 n/a 1307581 unspecified role unspecified role bait bait n/a 1307582 unspecified role unspecified role prey prey physical association physical association false false false Fig 4D erbb2-h2afy-2 1307575 n/a 1307581 unspecified role unspecified role prey prey n/a 1307582 unspecified role unspecified role bait bait physical association physical association false false false Fig 1C/D h2afy-erbb2-3 1307575 n/a 1307581 unspecified role unspecified role bait bait n/a 1307582 unspecified role unspecified role prey prey physical association physical association false false false Fig 4D h2afy-erbb2 1307576 n/a 1307582 unspecified role unspecified role prey prey n/a 1307583 unspecified role unspecified role bait bait c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false c_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag 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 3A Only mH2A1.2-MYC co-immunoprecipitated endogenous HER-2 (Fig. 3A). erbb2-h2afy-4 1307576 n/a 1307582 unspecified role unspecified role bait bait c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false c_terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1307583 unspecified role unspecified role prey prey halo_biotin_tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false macro_domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false physical association physical association false false false Fig 7 Only mH2A1.2-MYC co-immunoprecipitated endogenous HER-2 (Fig. 3A). erbb2-h2afy 1307577 n/a 1307581 unspecified role unspecified role prey prey n/a 1307582 unspecified role unspecified role bait bait n_terminus tandem tag tandem tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig 1A/B erbb2-h2afy-3 1307578 n/a 1307581 unspecified role unspecified role neutral component neutral component n/a 1307582 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig 4B h2afy-erbb2-4 1307579 n/a 1307583 unspecified role unspecified role bait bait n/a 1307584 unspecified role unspecified role prey prey promoter_region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig 6B At five of eight positions along the 5-kb stretch of the ERBB2 promoter, mH2A1.2 expression was significantly enriched. h2afy-erbb2-6 1307580 n/a 1307583 unspecified role unspecified role bait bait eis_domain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false O75367-3:p.[Glu202Gly;Ser204Ala] 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 region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1307582 unspecified role unspecified role prey prey physical association physical association false false false Fig 8G