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/ Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res. (0008-5472) 2007 imex curation Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org A mouse smooth muscle cDNA library was used. Only protein-protein interactions 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 Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org Accepted 2007-OCT-25 by JYOTI. A mouse smooth muscle cDNA library was used. Cancer Res. (0008-5472) 2007 Only protein-protein interactions imex curation Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res. (0008-5472) 2007 imex curation Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org A mouse smooth muscle cDNA library was used. Only protein-protein interactions human-ht1080 Homo sapiens HT1080 fribrosarcoma cell line ht1080 confirmed as human with IEF of AST, LDH, NP established from the biopsy from a fibrosarcoma of a 35-year-old Caucasian man in 1972 confocal microscopy confocal microscopy predetermined predetermined participant predetermined Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org Accepted 2007-OCT-25 by JYOTI. A mouse smooth muscle cDNA library was used. Cancer Res. (0008-5472) 2007 Only protein-protein interactions imex curation Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res. (0008-5472) 2007 imex curation Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org A mouse smooth muscle cDNA library was used. Only protein-protein interactions 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 Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org Accepted 2007-OCT-25 by JYOTI. A mouse smooth muscle cDNA library was used. Cancer Res. (0008-5472) 2007 Only protein-protein interactions imex curation Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Regulation of phosphoglucose isomerase/autocrine motility factor activities by the poly(ADP-ribose) polymerase family-14. Cancer Res. (0008-5472) 2007 imex curation Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org A mouse smooth muscle cDNA library was used. Only protein-protein interactions human-ht1080 Homo sapiens HT1080 fribrosarcoma cell line ht1080 confirmed as human with IEF of AST, LDH, NP established from the biopsy from a fibrosarcoma of a 35-year-old Caucasian man in 1972 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yanagawa T., Funasaka T., Tsutsumi S., Hu H., Watanabe H., Raz A. Cancer - Interactions investigated in the context of cancer The yeast two-hybrid assay was done using the CytoTrap system (Stratagene). Mouse pgi/amf gene was inserted into pSos vector to produce human son of sevenless (hSos) protein fused with mouse PGI/AMF, which was used as a bait protein. The cDNA library originated from mouse smooth muscle and was inserted into pMyr vector that contains DNA encoding the myristylation membrane localization signal (Myr), also purchased from Stratagene. raza@karmanos.org Accepted 2007-OCT-25 by JYOTI. A mouse smooth muscle cDNA library was used. Cancer Res. (0008-5472) 2007 Only protein-protein interactions imex curation par14_mouse Protein mono-ADP-ribosyltransferase PARP14 Parp14 Kiaa1268 Collaborator of STAT6 ADP-ribosyltransferase diphtheria toxin-like 8 Poly [ADP-ribose] polymerase 14 protein protein mouse Mus musculus Mouse MAASGSFPLLVEGSWGPDPPKNLINKLQVYFQSRKKSGGGECEVVPEPGNPARFRVLFSPEDVRQNVLERGNHELVWQEKGTFKLTVLMPTDPEEASASKKSRKESPEEESKTKEDAVKQGDLDITHSPSSGSEKTEDVPKECENISSMVAFENLPEKVSEMVLTILVENISGLPSDDFKVEVNRDFAVAVVTFQKPIDIKKFIVDCISHRSNQQLQLAPRLLETTNVVRVENLPPGVDEYQLQLFFENPFNGGGRVARVECFPEESSALVEFCDSKVLDTVMAKTHSYNKMPLSVFPYYPSLGTALYGEEKPLIKLPASFQESLDLPLWKFFQKNNHLIEEINNEMRCCHCELTWSEINGKLTIRPAATLVNHRLSIKTWQRDASAVLSGIKSKYGVELFEVCSPVWDIIKHELESGDDRVLVEFEKESLNIAGKSEDVQGMSQKIRELIESTTEKLRREEQSLKEKVAISPGKHYLLHHSGFLKDLSKGFPEMEISYDATAQFLYLKGFRADVYKVKCDIQEKVFSMAQKDVQVSSEVFEFLQQVDSQRLSKSLFEAQNILAIYELKGTALFLVGSSFKDLAEAETKMLSALSHKQIEVEDKEVLISNGWKKKVHPLQKRHSSCATIIVQNELTSETPAKVIVTGCVKEVNEIHRQLFEYLENNMKVERALKIKPSLIVDYLRTDKRLLSKIKKAHVYVHFKPKDNPNSILLTGCKSKVLECMNLVKEIQDSVCVQRFQTDKAGVRHFFKDKESYYKTEIGRQFGCVIELEEDREEKGEEEDGEEEEGEEEGESSINEQKCHLQRDIAPGVKLFVLEEDLSRFPVDVVVNAANENLKHISGLAQALSKAAGPELQTECDQIVKEGGVVLPGNAVISKAGKLPCHHVIHAVGPRWKGDKVLECVSLLKKVVRQSLSLAEEHRCRSIAMPAVSAGIFDFPLELCVANIVSAIKEHFQHKRDTHTLKKIYLVGLPAKVARAFAEAVKTTYKDSLSHTAFPSSLKALVPLGKTPQKQGSLLVSPEGLRIRLVEEGVQNATTHAIVNSISPDLKLNKGPLSQAFLEKAGPKLQEELTRSGQGVSVDVGTILQTSGCNLNSRHVFHVVPPPWKSNNSAWSLKIMKNIIRDCLKTTENLSLQSIAFPAIGTGNLRFPKPEFAKLIISEVLKFSSRNQLKTLQEVQFLLHPKDHENIQAFSDEFDKRNNGDPSDKNPKAEDTQGIYGSLSSPTLGMHEMNIGPILFQVATGNIIKEVADVIVNSTTLTFDLKSGVSKAILEGAGQNVEQECSLLAKQSNHGYIVTGGGLLQCKNIIHVVGGNDVKKSVSCVLEECEQRNYSSICLPAIGTGNAQQDPNVVAKAIIDAIEEFVQKKSVQAVKRVKVVIFQPHILQFFYDNMKEREGSPAPPKQSPAKQSVMSKIASFLGFPKQASPKKNTLVLEKKIEHTVFQVCGSGVDSVNKTISWLKELITKEQLSYTNDDECVSDFDMEEYEKLNEIQKELNITIEMNQKKTSIQVSGISRDVIKARDEIEGMIKSIRLAKEKESQADYISTYVEWQYIDKNITQCFDKMTNMKLEVAWKAKKKDTVVQIHNQDFTVDLSTNTATAPQGQTFTVQRLVKAEAEIPANWSDMKQDKLLLVNLQTSDPEYNMVASAFRQTCSNFFIEKIERIQNPALWRRYQAYKKSMDEKNGNVRNEKHLFHGTEASSLPQLNSNGFNRSYAGKNATAYGKGTYFAVKASYSACDTYSRPDTNGRKYMYYVRVLTGNYTNGNASLIVPPSRDPQNAADLYDTVTDNDKNPSIFVVFYDNQTYPEYLITFRQ DIP protein Q2EMV9 original sequence version: 82 42E902CC20858931 g6pi_mouse Glucose-6-phosphate isomerase Gpi1 Gpi Phosphoglucose isomerase Phosphohexose isomerase Neuroleukin Autocrine motility factor protein protein mouse Mus musculus Mouse MAALTRNPQFQKLLEWHRANSANLKLRELFEADPERFNNFSLNLNTNHGHILVDYSKNLVNKEVMQMLVELAKSRGVEAARDNMFSGSKINYTENRAVLHVALRNRSNTPIKVDGKDVMPEVNRVLDKMKSFCQRVRSGDWKGYTGKSITDIINIGIGGSDLGPLMVTEALKPYSKGGPRVWFVSNIDGTHIAKTLASLSPETSLFIIASKTFTTQETITNAETAKEWFLEAAKDPSAVAKHFVALSTNTAKVKEFGIDPQNMFEFWDWVGGRYSLWSAIGLSIALHVGFDHFEQLLSGAHWMDQHFLKTPLEKNAPVLLALLGIWYINCYGCETHALLPYDQYMHRFAAYFQQGDMESNGKYITKSGARVDHQTGPIVWGEPGTNGQHAFYQLIHQGTKMIPCDFLIPVQTQHPIRKGLHHKILLANFLAQTEALMKGKLPEEARKELQAAGKSPEDLEKLLPHKVFEGNRPTNSIVFTKLTPFILGALIAMYEHKIFVQGIMWDINSFDQWGVELGKQLAKKIEPELEGSSAVTSHDSSTNGLISFIKQQRDTKLE 7299E98B12B4C375 gpi-parp14 1 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association we screened a cDNA library using a yeast two-hybrid system to search for interacting protein(s) and report on the finding of poly(ADP-ribose) polymerase-14 (PARP-14) to be a binding partner with PGI/AMF. PREY gene XM_488522 The isolated prey plasmid is reported to differ from XM_488522 at the 4865th base pair and so at aa 1609. Note that the UniProtKB entry Q2EMV9 also differs from the translation of XM_488522 at aa 1609. The protein corresponding to the isolated prey plasmid is reported to be NH2-terminal truncated. gpi-parp14-1 3 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association PREY gene XM_488522 The isolated prey plasmid is reported to differ from XM_488522 at the 4865th base pair and so at aa 1609. Note that the UniProtKB entry Q2EMV9 also differs from the translation of XM_488522 at aa 1609. The protein corresponding to the prey plasmid is reported to be NH2-terminal truncated. 1 B gpi-parp14-2 4 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association 2 E HT-1080 cells prepared from monolayer cultures were immunoprecipitated with either anti-PGI/AMF polyclonal antibodies or preimmunized rabbit IgG and blotted with anti-PARP-14 or anti-PGI/AMF antibodies (Fig. 2E). The results clearly show that PARP-14 coprecipitated with PGI/AMF, suggesting that the two are complexed in vivo. gpi-parp14-3 2 5 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 C we confirmed that PARP-14-PGI/AMF are indeed colocalized in vivo in HT-1080 cells by immunofluorescence analysis (Fig. 3) using confocal microscopy. PARP-14 was found to predominantly localize at the perinuclear region and slightly at the peripheral region of the cells (Fig. 3A), whereas PGI/AMF was distributed throughout the cytoplasm (Fig. 3B) and colocalization was readily observed mainly in the perinuclear region