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/ Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Genes Dev. (0890-9369) 1998 imex curation Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de 14-May-2008: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de Accepted 2008-APR-28 by CDEROW. 14-May-2008: Contacted by JYOTI. Genes Dev. (0890-9369) 1998 Only protein-protein interactions imex curation Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Genes Dev. (0890-9369) 1998 imex curation Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de 14-May-2008: Contacted by JYOTI. 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 Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis Its not clear to me if the n-terminal construct interacted with the prey. The bait has no range now. koncz@mpiz-koeln.mpg.de Accepted 2008-APR-28 by CDEROW. 14-May-2008: Contacted by JYOTI. Arabidopsis cell suspension cDNA library. Genes Dev. (0890-9369) 1998 Only protein-protein interactions imex curation Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Genes Dev. (0890-9369) 1998 imex curation Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de 14-May-2008: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de Accepted 2008-APR-28 by CDEROW. 14-May-2008: Contacted by JYOTI. Genes Dev. (0890-9369) 1998 Only protein-protein interactions imex curation Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Pleiotropic control of glucose and hormone responses by PRL1, a nuclear WD protein, in Arabidopsis. Genes Dev. (0890-9369) 1998 imex curation Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de 14-May-2008: Contacted by JYOTI. 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 Nemeth K., Salchert K., Putnoky P., Bhalerao R., Koncz-Kalman Z., Stankovic-Stangeland B., Bako L., Mathur J., Okresz L., Stabel S., Geigenberger P., Stitt M., Redei GP., Schell J., Koncz C. Apoptosis - Interactions involving proteins with a function related to apoptosis koncz@mpiz-koeln.mpg.de Accepted 2008-APR-28 by CDEROW. 14-May-2008: Contacted by JYOTI. Genes Dev. (0890-9369) 1998 Only protein-protein interactions imex curation kpcb_human Protein kinase C beta type PRKCB1 PKCB PRKCB protein protein human Homo sapiens Human MADPAAGPPPSEGEESTVRFARKGALRQKNVHEVKNHKFTARFFKQPTFCSHCTDFIWGFGKQGFQCQVCCFVVHKRCHEFVTFSCPGADKGPASDDPRSKHKFKIHTYSSPTFCDHCGSLLYGLIHQGMKCDTCMMNVHKRCVMNVPSLCGTDHTERRGRIYIQAHIDRDVLIVLVRDAKNLVPMDPNGLSDPYVKLKLIPDPKSESKQKTKTIKCSLNPEWNETFRFQLKESDKDRRLSVEIWDWDLTSRNDFMGSLSFGISELQKASVDGWFKLLSQEEGEYFNVPVPPEGSEANEELRQKFERAKISQGTKVPEEKTTNTVSKFDNNGNRDRMKLTDFNFLMVLGKGSFGKVMLSERKGTDELYAVKILKKDVVIQDDDVECTMVEKRVLALPGKPPFLTQLHSCFQTMDRLYFVMEYVNGGDLMYHIQQVGRFKEPHAVFYAAEIAIGLFFLQSKGIIYRDLKLDNVMLDSEGHIKIADFGMCKENIWDGVTTKTFCGTPDYIAPEIIAYQPYGKSVDWWAFGVLLYEMLAGQAPFEGEDEDELFQSIMEHNVAYPKSMSKEAVAICKGLMTKHPGKRLGCGPEGERDIKEHAFFRYIDWEKLERKEIQPPYKPKARDKRDTSNFDKEFTRQPVELTPTDKLFIMNLDQNEFAGFSYTNPEFVINV 3937E7B667108C68 prl1_arath Protein pleiotropic regulatory locus 1 PRL1 At4g15900 dl3990w MAC2 MOS4-associated complex protein 2 FCAALL.40 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPAPTTEIEPIEAQSLKKLSLKSLKRSLELFSPVHGQFPPPDPEAKQIRLSHKMKVAFGGVEPVVSQPPRQPDRINEQPGPSNALSLAAPEGSKSTQKGATESAIVVGPTLLRPILPKGLNYTGSSGKSTTIIPANVSSYQRNLSTAALMERIPSRWPRPEWHAPWKNYRVIQGHLGWVRSVAFDPSNEWFCTGSADRTIKIWDVATGVLKLTLTGHIEQVRGLAVSNRHTYMFSAGDDKQVKCWDLEQNKVIRSYHGHLSGVYCLALHPTLDVLLTGGRDSVCRVWDIRTKMQIFALSGHDNTVCSVFTRPTDPQVVTGSHDTTIKFWDLRYGKTMSTLTHHKKSVRAMTLHPKENAFASASADNTKKFSLPKGEFCHNMLSQQKTIINAMAVNEDGVMVTGGDNGSIWFWDWKSGHSFQQSETIVQPGSLESEAGIYAACYDNTGSRLVTCEADKTIKMWKEDENATPETHPINFKPPKEIRRF 604D3E6FCDA8A998 impa3_arath Importin subunit alpha-3 IMPA3 KAP2 Karyopherin subunit alpha-2 MOS6 Protein MODIFIER OF SNC1 6 T10M13.16 At4g02150 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSLRPSAKTEVRRNRYKVAVDAEEGRRRREDNLVEIRKNKREENLQKKRFTSSMAFGSATGQTEQDLSSANQLKDNLPAMVAGIWSEDSNSQLEATNLLRKLLSIEQNPPINEVVQSGVVPRVVKFLSRDDFPKLQFEAAWALTNIASGTSENTNVIIESGAVPIFIQLLSSASEDVREQAVWALGNVAGDSPKCRDLVLSYGAMTPLLSQFNENTKLSMLRNATWTLSNFCRGKPPPAFEQTQPALPVLERLVQSMDEEVLTDACWALSYLSDNSNDKIQAVIEAGVVPRLIQLLGHSSPSVLIPALRTIGNIVTGDDLQTQMVLDQQALPCLLNLLKNNYKKSIKKEACWTISNITAGNADQIQAVIDAGIIQSLVWVLQSAEFEVKKEAAWGISNATSGGTHDQIKFMVSQGCIKPLCDLLTCPDLKVVTVCLEALENILVVGEAEKNLGHTGEDNLYAQMIDEAEGLEKIENLQSHDNNDIYDKAVKILETFWTEDNEEEGNDENHAPQSGFQFGSTNVPPGQFNFI 55B8979D205FCAA3 prl1-pkcb 1 5 unspecified role unspecified role prey prey spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. 6 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4e Following stringent washes, immunoblotting of the matrix-bound proteins revealed that GST-PRL1-Delta B retained PKC-beta II, but not PKC-beta I, The authors stated that "Because PKC-beta I and beta II sequences only differed by 52 carboxy-terminal amino acids (Kubo et al. 1987), the data also indicated that the carboxyl terminus of PKC-beta II was required for PRL1-binding". The expressed in annotation for the P05771 protein is spofr-sf_21, although the authors only state the protein was expressed in an insect based protein expression system. I have assumed this is based on the spofr-sf_21 cell expression system. prl1-kap2 2 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association To screen for Arabidopsis cDNAs encoding PRL1-interacting proteins (PIPs), the full-length PRL1 protein and an amino-terminal PRL1 segment of 321 amino acids were expressed as baits carrying the Gal4p DNA-binding domain in yeast using the two-hybrid vector pAS2 (Durfee et al. 1993). From 18.4 x 107 transformants obtained with a cDNA expression library prepared from an Arabidopsis cell suspension in pACT2 (Durfee et al. 1993), 342 clones showed His+ and LacZ+ phenotype indicating an interaction between the PRL1 baits and cDNA encoded proteins fused to the activation domain in pACT2 (Fig. 7a). Classification of PIP clones identified a family of 62 cDNAs coding for carboxy-terminal segments of PIP-B corresponding to a novel class of Arabidopsis alpha -importins, ATHKAP2. prl1-kap2-1 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 6 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 7b The His- phenotype of yeast strains carrying an ATHKAP2-fused activation domain in combination with the Gal4p DNA-binding domain either alone or in fusion with unrelated proteins (NPK5, PIP-M, and PIP-N; Fig. 7b) showed that the PRL1-ATHKAP2 interaction was specific. prl1-kap2-2 3 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 7 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 7c To support these data, [35S]methionine-labeled PRL1 was synthesized by coupled transcription and translation using the cDNA template, and equal aliquots of PRL1 protein were incubated with GST-ATHKAP2 and GST proteins immobilized on glutathione-S-Sepharose, as well as with the empty Sepharose matrix (see Materials and Methods). PRL1 was quantitatively removed from the control Sepharose and GST matrices by stringent washes, but remained tightly-bound to GST-ATHKAP2, confirming an interaction of ATHKAP2 with PRL1 in vitro