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/ Identification and characterization of follistatin as a novel angiogenin-binding protein. Identification and characterization of follistatin as a novel angiogenin-binding protein. FEBS Lett. (0014-5793) 2007 imex curation Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. davet 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 Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn Accepted 2008-FEB-16 by DAVET. 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. FEBS Lett. (0014-5793) 2007 davet Only protein-protein interactions imex curation Identification and characterization of follistatin as a novel angiogenin-binding protein. Identification and characterization of follistatin as a novel angiogenin-binding protein. FEBS Lett. (0014-5793) 2007 imex curation Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn Accepted 2008-FEB-16 by DAVET. 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. FEBS Lett. (0014-5793) 2007 davet Only protein-protein interactions imex curation Identification and characterization of follistatin as a novel angiogenin-binding protein. Identification and characterization of follistatin as a novel angiogenin-binding protein. FEBS Lett. (0014-5793) 2007 imex curation Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn Accepted 2008-FEB-16 by DAVET. 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. FEBS Lett. (0014-5793) 2007 davet Only protein-protein interactions imex curation Identification and characterization of follistatin as a novel angiogenin-binding protein. Identification and characterization of follistatin as a novel angiogenin-binding protein. FEBS Lett. (0014-5793) 2007 imex curation Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. davet 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 Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 Source of angiogenin from PMID 11549292 Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn Accepted 2008-FEB-16 by DAVET. 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. FEBS Lett. (0014-5793) 2007 davet Only protein-protein interactions imex curation Identification and characterization of follistatin as a novel angiogenin-binding protein. Identification and characterization of follistatin as a novel angiogenin-binding protein. FEBS Lett. (0014-5793) 2007 imex curation Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Gao X., Hu H., Zhu J., Xu Z. Cancer - Interactions investigated in the context of cancer Source of angiogenin from PMID 11549292 zpxu@zju.edu.cn Accepted 2008-FEB-16 by DAVET. 17-April-2008: Contacted by JYOTI. A human heart cDNA library was used. FEBS Lett. (0014-5793) 2007 davet Only protein-protein interactions imex curation fst_human Follistatin Activin-binding protein FST protein protein human Homo sapiens Human MVRARHQPGGLCLLLLLLCQFMEDRSAQAGNCWLRQAKNGRCQVLYKTELSKEECCSTGRLSTSWTEEDVNDNTLFKWMIFNGGAPNCIPCKETCENVDCGPGKKCRMNKKNKPRCVCAPDCSNITWKGPVCGLDGKTYRNECALLKARCKEQPELEVQYQGRCKKTCRDVFCPGSSTCVVDQTNNAYCVTCNRICPEPASSEQYLCGNDGVTYSSACHLRKATCLLGRSIGLAYEGKCIKAKSCEDIQCTGGKKCLWDFKVGRGRCSLCDELCPDSKSDEPVCASDNATYASECAMKEAACSSGVLLEVKHSGSCNSISEDTEEEEEDEDQDYSFPISSILEW D9BB45055D84AC90 p19883-2 Follistatin FST Activin-binding protein FS288 FS-288 protein protein human Homo sapiens Human MVRARHQPGGLCLLLLLLCQFMEDRSAQAGNCWLRQAKNGRCQVLYKTELSKEECCSTGRLSTSWTEEDVNDNTLFKWMIFNGGAPNCIPCKETCENVDCGPGKKCRMNKKNKPRCVCAPDCSNITWKGPVCGLDGKTYRNECALLKARCKEQPELEVQYQGRCKKTCRDVFCPGSSTCVVDQTNNAYCVTCNRICPEPASSEQYLCGNDGVTYSSACHLRKATCLLGRSIGLAYEGKCIKAKSCEDIQCTGGKKCLWDFKVGRGRCSLCDELCPDSKSDEPVCASDNATYASECAMKEAACSSGVLLEVKHSGSCN B516643224C7F8FF angi_human Angiogenin Ribonuclease 5 ANG RNASE5 protein protein human Homo sapiens Human MVMGLGVLLLVFVLGLGLTPPTLAQDNSRYTHFLTQHYDAKPQGRDDRYCESIMRRRGLTSPCKDINTFIHGNKRSIKAICENKNGNPHRENLRISKSSFQVTTCKLHGGSPWPPCQYRATAGFRNVVVACENGLPVHLDQSIFRRP 9C462DA3C8D39ACC ang-fst 4 7 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait physical association physical association follistatin was identified as a binding partner of angiogenin by a yeast two-hybrid screen PREY gene NM_006350 Fig. 4 A ang-fst-1 1 7 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait physical association physical association author score author-based confidence author score PREY gene NM_006350 To reconfirm the interaction between angiogenin and follistatin, yeast cells were co-transformed with pDBLeu or pDBLeu-ANG along with pPC86 or pPC86-FS. Transformants turned blue in X-gal analysis only when angiogenin and follistatin were simultaneously expressed Fig. 1 A, 4 fst-ang 3 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 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 ecobb Escherichia coli (strain B / BL21) physical association physical association 1 B Angiogenin was pulled down by follistatin specifically. ang-fst-2 2 8 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Fig. 2 C FS co-localizes with ANG in the nucleus ang-fst-3 5 8 unspecified role unspecified role fluorescence donor fluorescence donor region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Fig. 3 FRET analysis demonstrates that angiogenin interacts with follistatin in HeLa cell nucleus.