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/ Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 imex curation Bhalerao RP., Salchert K., Bako L., Okresz L., Szabados L., Muranaka T., Machida Y., Schell J., Koncz C. It was not possible to obtain enough details of the thioredoxin construct used in the kinase assays to allow curation of these assays. koncz@mpiz-koeln.mpg.de 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. 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 Bhalerao RP., Salchert K., Bako L., Okresz L., Szabados L., Muranaka T., Machida Y., Schell J., Koncz C. It was not possible to obtain enough details of the thioredoxin construct used in the kinase assays to allow curation of these assays. koncz@mpiz-koeln.mpg.de Accepted 2008-Not a month!-29 by JYOTI. 03-Mar-2009: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 jyoti Only protein-protein interactions imex curation Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 imex curation Bhalerao RP., Salchert K., Bako L., Okresz L., Szabados L., Muranaka T., Machida Y., Schell J., Koncz C. It was not possible to obtain enough details of the thioredoxin construct used in the kinase assays to allow curation of these assays. koncz@mpiz-koeln.mpg.de 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Bhalerao RP., Salchert K., Bako L., Okresz L., Szabados L., Muranaka T., Machida Y., Schell J., Koncz C. It was not possible to obtain enough details of the thioredoxin construct used in the kinase assays to allow curation of these assays. koncz@mpiz-koeln.mpg.de Accepted 2008-Not a month!-29 by JYOTI. 03-Mar-2009: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 1999 jyoti Only protein-protein interactions imex curation aakg_yeast 5'-AMP-activated protein kinase subunit gamma SNF4 CAT3 Regulatory protein CAT3 SCI1 Sucrose non-fermenting protein 4 YGL115W protein protein yeast Saccharomyces cerevisiae Baker's yeast MKPTQDSQEKVSIEQQLAVESIRKFLNSKTSYDVLPVSYRLIVLDTSLLVKKSLNVLLQNSIVSAPLWDSKTSRFAGLLTTTDFINVIQYYFSNPDKFELVDKLQLDGLKDIERALGVDQLDTASIHPSRPLFEACLKMLESRSGRIPLIDQDEETHREIVVSVLTQYRILKFVALNCRETHFLKIPIGDLNIITQDNMKSCQMTTPVIDVIQMLTQGRVSSVPIIDENGYLINVYEAYDVLGLIKGGIYNDLSLSVGEALMRRSDDFEGVYTCTKNDKLSTIMDNIRKARVHRFFVVDDVGRLVGVLTLSDILKYILLGSN 51B387E346EE9561 51B387E346EE9561 kin11_arath SNF1-related protein kinase catalytic subunit alpha KIN11 KIN11 AKIN alpha-1 AKIN11 SNF1-related kinase 1.2 SNR1 SNRK1.2 MXE2.16 At3g29160 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDHSSNRFGNNGVESILPNYKLGKTLGIGSFGKVKIAEHVVTGHKVAIKILNRRKIKNMEMEEKVRREIKILRLFMHPHIIRQYEVIETTSDIYVVMEYVKSGELFDYIVEKGRLQEDEARNFFQQIISGVEYCHRNMVVHRDLKPENLLLDSRCNIKIADFGLSNVMRDGHFLKTSCGSPNYAAPEVISGKLYAGPEVDVWSCGVILYALLCGTLPFDDENIPNLFKKIKGGIYTLPSHLSSEARDLIPRMLIVDPVKRITIPEIRQHRWFQTHLPRYLAVSPPDTVEQAKKINEEIVQEVVNMGFDRNQVLESLRNRTQNDATVTYYLLLDNRFRVPSGYLESEFQETTDSGSNPMRTPEAGASPVGHWIPAHVDHYGLGARSQVPVDRKWALGLQSHAHPREIMNEVLKALQELNVCWKKIGHYNMKCRWVPGLADGQNTMVNNQLHFRDESSIIEDDCAMTSPTVIKFELQLYKAREEKYLLDIQRVNGPQFLFLDLCAAFLTELRVI 4FBEF55D3EC60F87 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 q38997-2 SNF1-related protein kinase catalytic subunit alpha KIN10 KIN10 AK21 AKIN10 SKIN10 SNR2 SNRK1.1 T4P13.22 At3g01090 AKIN alpha-2 SNF1-related kinase 1.1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDGSGTGSRSGVESILPNYKLGRTLGIGSFGRVKIAEHALTGHKVAIKILNRRKIKNMEMEEKVRREIKILRLFMHPHIIRLYEVIETPTDIYLVMEYVNSGELFDYIVEKGRLQEDEARNFFQQIISGVEYCHRNMVVHRDLKPENLLLDSKCNVKIADFGLSNIMRDGHFLKTSCGSPNYAAPEVISGKLYAGPEVDVWSCGVILYALLCGTLPFDDENIPNLFKKIKGGIYTLPSHLSPGARDLIPRMLVVDPMKRVTIPEIRQHPWFQAHLPRYLAVPPPDTVQQAKKIDEEILQEVINMGFDRNHLIESLRNRTQNDGTVTYYLILDNRFRASSGYLGAEFQETMEGTPRMHPAESVASPVSHRLPGLMEYQGVGLRSQYPVERKWALGLQSRAHPREIMTEVLKALQDLNVCWKKIGHYNMKCRWVPNSSADGMLSNSMHDNNYFGDESSIIENEAAVKSPNVVKFEIQLYKTRDDKYLLDLQRVQGPQFLFLDLCAAFLAQLRVL 5A18655A0AA506DF snf1_yeast Carbon catabolite-derepressing protein kinase SNF1 CAT1 CCR1 GLC2 PAS14 Sucrose nonfermentating protein 1 D8035.20 YDR477W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSNNNTNTAPANANSSHHHHHHHHHHHHHGHGGSNSTLNNPKSSLADGAHIGNYQIVKTLGEGSFGKVKLAYHTTTGQKVALKIINKKVLAKSDMQGRIEREISYLRLLRHPHIIKLYDVIKSKDEIIMVIEYAGNELFDYIVQRDKMSEQEARRFFQQIISAVEYCHRHKIVHRDLKPENLLLDEHLNVKIADFGLSNIMTDGNFLKTSCGSPNYAAPEVISGKLYAGPEVDVWSCGVILYVMLCRRLPFDDESIPVLFKNISNGVYTLPKFLSPGAAGLIKRMLIVNPLNRISIHEIMQDDWFKVDLPEYLLPPDLKPHPEEENENNDSKKDGSSPDNDEIDDNLVNILSSTMGYEKDEIYESLESSEDTPAFNEIRDAYMLIKENKSLIKDMKANKSVSDELDTFLSQSPPTFQQQSKSHQKSQVDHETAKQHARRMASAITQQRTYHQSPFMDQYKEEDSTVSILPTSLPQIHRANMLAQGSPAASKISPLVTKKSKTRWHFGIRSRSYPLDVMGEIYIALKNLGAEWAKPSEEDLWTIKLRWKYDIGNKTNTNEKIPDLMKMVIQLFQIETNNYLVDFKFDGWESSYGDDTTVSNISEDEMSTFSAYPFLHLTTKLIMELAVNSQSN F5C63565C986C4E3 F5C63565C986C4E3 y5872_arath SMR domain-containing protein At5g58720 PIPC PRL1-interacting protein PIPC MZN1.16 At5g58720 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MKQKNQHKKKKKRSCAAKPSGDGTTSDGNKKDVEEERKDGEGKREIENVGKNFIESLMEAFCSVSMEEAMAAYKEAGGDLNKAAEILSDLVESGDDPSTSSVASGSSGQETASTSEYGAGSSSSCSEDLTRDRWFKGSKQSRVIAATGMVSSVIAKDYLKPNPVRKEFPMMERSKELCGNGKKAADREKAEQFLSSMLGDDCELSMAVVRDVLCQCGYDVDMALNVLLDMSSSSTDDSLSGKCFGIGVSDSLAESSFDTDTSDCELFWGGDYSQRDYAKALMSSQDPFATTQGIDELGLPQKVLESLFNIRQNPKHESKTTSWRNVAKKMQSLGIDASSSSGEEPHPNTFVKDDSYHELRKGANDQWNVTKSYYQKAAEAYSKGGRAHAAYLSDKGRVASKQAQRADERASQDIFVARNKGIENVVTIDLHGQHVKPAMKLLKLHLLFGSYVPSIQTLRVITGCGASGFGKSKVKQSVVKLLEREGVRYCEENRGTLLIKLDGGSREFSFLDTESDSDE 1220DB469C4D87F8 snf1-prl1 1 7 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey n-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 1B The yeast strain expressing GBD-SNF1 and GADPRL1 grew in the presence of 50 mM 3-aminotriazole HIS3 inhibitor, as well as the control, and displayed LacZ1 phenotype in filter lift assays (16, 17), indicating that an interaction between SNF1p and PRL1 led to the activation of the GAL4- controlled HIS3 and LacZ reporter genes. A similar assay with pACT2prl11-620 and pAS1snf1 showed that PRL1 sequences located between the N terminus and amino acid position 195 were involved in the binding of SNF1p The authors concluded that the "assay with pACT2prl11-620 and pAS1snf1 showed that PRL1 sequences located between the N terminus and amino acid position 195 were involved in the binding of SNF1p". snf1-snf4-7 1 3 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 1B prl1-kin10 1 5 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 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 1 Interactions of AKIN10 and AKIN11 with SNF4p and PRL1 in the yeast two-hybrid system kin10-snf4 1 3 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 1 Figure 1D The GBD-fusion protein carrying the AKIN10 catalytic domain between positions 1 and 381 did not interact either with GAD-PRL11�195, carrying 195 N-terminal amino acids from PRL1, or GAD-SNF4p 2% glucose (weight/volume) kin11-snf4 1 3 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association Table 1 Figure 1D Interactions of AKIN10 and AKIN11 with SNF4p and PRL1 in the yeast two-hybrid system 2% glucose (weight/volume) kin10-prl1 2 6 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 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 direct interaction direct interaction Figure 1E To confirm that no yeast proteins were involved as mediators in the PRL1-AKIN interactions observed in the two-hybrid system, N-terminal GST fusions with AKIN10 and AKIN11 were constructed, purified, and immobilized on glutathione- Sepharose. [35S]Methione-labeled PRL1 protein was synthesized by coupled transcription�translation by using the cDNA as tem-plate and incubated with GST-AKIN10 and GST-AKIN11, as well as with control GST, and glutathione-Sepharose matrices. After stringent washes, the supernatant fractions and matrixbound proteins were resolved by SDSyPAGE and detected by autoradiography, demonstrating specific binding of PRL1 to both GST-AKIN10 and GST-AKIN11 in vitro kin11-prl1 2 5 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 4 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro direct interaction direct interaction Figure 1E To confirm that no yeast proteins were involved as mediators in the PRL1-AKIN interactions observed in the two-hybrid system, N-terminal GST fusions with AKIN10 and AKIN11 were constructed, purified, and immobilized on glutathione- Sepharose. [35S]Methione-labeled PRL1 protein was synthesized by coupled transcription�translation by using the cDNA as tem-plate and incubated with GST-AKIN10 and GST-AKIN11, as well as with control GST, and glutathione-Sepharose matrices. After stringent washes, the supernatant fractions and matrixbound proteins were resolved by SDSyPAGE and detected by autoradiography, demonstrating specific binding of PRL1 to both GST-AKIN10 and GST-AKIN11 in vitro kin10-prl1-1 1 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 5 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Table 1 Figure 1C Figure 1D Interactions of AKIN10 and AKIN11 with SNF4p and PRL1 in the yeast two-hybrid system 2% glucose (weight/volume) kin11-prl1-1 1 5 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role bait bait physical association physical association Figure 1C Figure 1D Interactions of AKIN10 and AKIN11 with SNF4p and PRL1 in the yeast two-hybrid system 2% glucose (weight/volume) prl1-pipc 1 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association Figure 1D 2% glucose (weight/volume)