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/ Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. J. Gen. Virol. (0022-1317) 2006 imex curation Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes 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 Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes was used. J. Gen. Virol. (0022-1317) 2006 Only protein-protein interactions imex curation Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. J. Gen. Virol. (0022-1317) 2006 imex curation Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes was used. Only protein-protein interactions human-rhek_1 Homo sapiens keratinocyte cell line RHEK-1 rhek_1 human keratinocyte cell line RHEK-1 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Gen. Virol. (0022-1317) 2006 Only protein-protein interactions imex curation Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. J. Gen. Virol. (0022-1317) 2006 imex curation Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes was used. Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Gen. Virol. (0022-1317) 2006 Only protein-protein interactions imex curation Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. J. Gen. Virol. (0022-1317) 2006 imex curation Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes 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 Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Gen. Virol. (0022-1317) 2006 Only protein-protein interactions imex curation Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. Human cellular protein VRK2 interacts specifically with Epstein-Barr virus BHRF1, a homologue of Bcl-2, and enhances cell survival. J. Gen. Virol. (0022-1317) 2006 imex curation Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw 12-MAR-2007: Contacted by JYOTI. A cDNA library from human normal foreskin keratinocytes was used. Only protein-protein interactions human-rhek_1 Homo sapiens keratinocyte cell line RHEK-1 rhek_1 human keratinocyte cell line RHEK-1 confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Li LY., Liu MY., Shih HM., Tsai CH., Chen JY. Apoptosis - Interactions involving proteins with a function related to apoptosis The cells used were a stably-transfected RHEK-1 cell line, NR27-2, characterized by doxycycline inducible BHFR1 expression. myliu@mail1.ntcn.edu.tw,cjy@nhri.org.tw Accepted 2007-APR-1 by JYOTI. 12-MAR-2007: Contacted by JYOTI. J. Gen. Virol. (0022-1317) 2006 Only protein-protein interactions imex curation ear_ebvb9 Apoptosis regulator BHRF1 BHRF1 Nuclear antigen Early antigen protein R protein protein ebvb9 Epstein-Barr virus (strain B95-8) (HHV-4) Human herpesvirus 4 HHV-4 MAYSTREILLALCIRDSRVHGNGTLHPVLELAARETPLRLSPEDTVVLRYHVLLEEIIERNSETFTETWNRFITHTEHVDLDFNSVFLEIFHRGDPSLGRALAWMAWCMHACRTLCCNQSTPYYVVDLSVRGMLEASEGLDGWIHQQGGWSTLIEDNIPGSRRFSWTLFLAGLTLSLLVICSYLFISRGRH DIP protein P03182 original sequence version: 91 8108BCB94F81DC8B bcl2_human Apoptosis regulator Bcl-2 BCL2 protein protein human Homo sapiens Human MAHAGRTGYDNREIVMKYIHYKLSQRGYEWDAGDVGAAPPGAAPAPGIFSSQPGHTPHPAASRDPVARTSPLQTPAAPGAAAGPALSPVPPVVHLTLRQAGDDFSRRYRRDFAEMSSQLHLTPFTARGRFATVVEELFRDGVNWGRIVAFFEFGGVMCVESVNREMSPLVDNIALWMTEYLNRHLHTWIQDNGGWDAFVELYGPSMRPLFDFSWLSLKTLLSLALVGACITLGAYLGHK DIP protein P10415 original sequence version: 148 DIP protein P10415 original sequence version: 125 DIP protein P10415 original sequence version: 151 DIP protein P10415 original sequence version: 204 DIP protein P10415 original sequence version: 149 3C49F2B714DC9CCB vrk2_human Serine/threonine-protein kinase VRK2 VRK2 Vaccinia-related kinase 2 protein protein human Homo sapiens Human MPPKRNEKYKLPIPFPEGKVLDDMEGNQWVLGKKIGSGGFGLIYLAFPTNKPEKDARHVVKVEYQENGPLFSELKFYQRVAKKDCIKKWIERKQLDYLGIPLFYGSGLTEFKGRSYRFMVMERLGIDLQKISGQNGTFKKSTVLQLGIRMLDVLEYIHENEYVHGDIKAANLLLGYKNPDQVYLADYGLSYRYCPNGNHKQYQENPRKGHNGTIEFTSLDAHKGVALSRRSDVEILGYCMLRWLCGKLPWEQNLKDPVAVQTAKTNLLDELPQSVLKWAPSGSSCCEIAQFLVCAHSLAYDEKPNYQALKKILNPHGIPLGPLDFSTKGQSINVHTPNSQKVDSQKAATKQVNKAHNRLIEKKVHSERSAESCATWKVQKEEKLIGLMNNEAAQESTRRRQKYQESQEPLNEVNSFPQKISYTQFPNSFYEPHQDFTSPDIFKKSRSPSWYKYTSTVSTGITDLESSTGLWPTISQFTLSEETNADVYYYRIIIPVLLMLVFLALFFL 9F3F6FCC9280568F bhfr1-vrk2 1 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Several clones were identified to interact with BHRF1 specifically and reproducibly. One of them contains cDNA sequence corresponding to the coding sequence of the VRK2 protein The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. bhfr1-vrk2-1 4 8 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 6 unspecified role unspecified role bait bait physical association physical association 1 C, 5 B, C, D The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. The two hybrid interaction assays showed that co-transformation of cells with BHRF1 and VRK2 resulted in activation of the HIS3 and lacZ reporter genes, as indicated by growth of the transformants on an SC-Trp, Ura, Leu, Lys, His plate (a His2 plate) and the formation of blue colonies on the colony-lift filter with a b-galactosidase assay. The deletion analysis suggested the last 108 C-terminal residues of VRK2 are important for the interaction. However, as VRK2-(94-508) did not interact with BHRF1, it may be that the first 93 aa contribute to a conformation required for interaction, or there may be an inhibitory domain in aa 94-174. bhfr1-vrk2-2 3 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 C The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. HA-tagged VRK2 could be pulled down specifically by immobilized GST-BHRF1, but not by GST alone bhfr1-vrk2-3 5 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 3 The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. HA-tagged VRK2 appeared to have a pattern of cytoplasmic staining in the transfected cells that was virtually identical to that of BHRF1. Q86Y07; P03182 transfection vrk2-bhfr1 2 8 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physical association physical association 4 B The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. Q86Y07; P03182 transfection From the results shown in Fig. 4(b), BHRF1 could be precipitated from HA-tagged VRK2-transfected N27-2 cells by using anti-HA antibody. bhfr1-vrk2-5 2 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait physical association physical association 4 B The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. Q86Y07; P03182 transfection From the results shown in Fig. 4(b), BHRF1 could be precipitated from HA-tagged VRK2-transfected N27-2 cells by using anti-HA antibody. The interaction was also observed in a reciprocal experiment This interaction is shown in 4 B, although in the text there is a note that data is not shown. bhfr1-vrk2-6 4 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 1 C, 5 F, G, H The protein P03182 relates to BHRF1 from EBV strain B95-8, but the authors in the paper Li et al., 2001 referred to do not specify the strain of EBV. This is the only UniProtKb entry corresponding to BHFR1. The length of the protein used and P03182 are the same. The two hybrid interaction assays showed that co-transformation of cells with BHRF1 and VRK2 resulted in activation of the HIS3 and lacZ reporter genes, as indicated by growth of the transformants on an SC-Trp, Ura, Leu, Lys, His plate (a His2 plate) and the formation of blue colonies on the colony-lift filter with a b-galactosidase assay. Deletion analysis suggests the first 18 aa (including BH4 domain) and aa 89-142 (including BH1 domain) of BHRF1 are important for binding VRK2. bcl2-vrk2 4 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 1 C, 5 F, G, H There was a positive reaction after 8 h. Thus the authors term this weaker than the interaction with BHFR1 which occurs within 30 min. whereas formation of faint blue colonies was not observed within the first 8 h when BHRF1 was replaced by Bcl-2.