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/ Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell (1040-4651) 2001 imex curation De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. diinz@gengenp.rug.ac.be 25-Jun-2008: Contacted by JYOTI. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) orchard 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 De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. diinz@gengenp.rug.ac.be Accepted 2008-JUL-10 by ORCHARD. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) Plant Cell (1040-4651) 2001 orchard Only protein-protein interactions imex curation Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell (1040-4651) 2001 imex curation De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. diinz@gengenp.rug.ac.be 25-Jun-2008: Contacted by JYOTI. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) orchard Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. Note the MM states that the PCR product for KRP3 started at Met 11 Protein was extracted from a 2 day old Arabidopsis cell suspension culture. The KRP3 protein was coupled to cyanogen bromide-activated Sepharose 4B (Amersham Pharmacia Biotech) at a concentration of 5 mg/mL according to the manufacturer's instructions. diinz@gengenp.rug.ac.be Accepted 2008-JUL-10 by ORCHARD. 25-Jun-2008: Contacted by JYOTI. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) Plant Cell (1040-4651) 2001 orchard Only protein-protein interactions imex curation Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell (1040-4651) 2001 imex curation De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. diinz@gengenp.rug.ac.be 25-Jun-2008: Contacted by JYOTI. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) orchard 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 De Veylder L., Beeckman T., Beemster GT., Krols L., Terras F., Landrieu I., van der Schueren E., Maes S., Naudts M., Inze D. diinz@gengenp.rug.ac.be Accepted 2008-JUL-10 by ORCHARD. 25-Jun-2008: Contacted by JYOTI. A Arabidopsis flower cDNA library obtained from the Arabidopsis Biological Resources Center (library number CD4-6; Columbus, OH) Plant Cell (1040-4651) 2001 orchard Only protein-protein interactions imex curation krp6_arath Cyclin-dependent kinase inhibitor 6 KRP6 ACK1 At3g19150 ICK4 MVI11.5 KIP-related protein 6 Inhibitor/interactor of CDK protein 4 Cyclin-dependent kinase inhibitor p22ack1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSERKRELAEEASSTSFSPLKKTKLNDSSDSSPDSHDVIVFAVSSSSVASSAALASDECSVTIGGEESDQSSSISSGCFTSESKEIAKNSSSFGVDLEDHQIETETETSTFITSNFRKETSPVSEGLGETTTEMESSSATKRKQPGVRKTPTAAEIEDLFSELESQDDKKKQFIEKYNFDIVNDEPLEGRYKWDRL DIP protein Q0WNX9 original sequence version: 53 AC543304CE6B92C4 ckb11_arath Cyclin-dependent kinase B1-1 CDKB1-1 At3g54180 F24B22.140 CDC2B Cell division control protein 2 homolog B protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEKYEKLEKVGEGTYGKVYKAMEKGTGKLVALKKTRLEMDEEGIPPTALREISLLQMLSTSIYVVRLLCVEHVHQPSTKSQSTKSNLYLVFEYLDTDLKKFIDSYRKGPNPKPLEPFLIQKLMFQLCKGVAHCHSHGVLHRDLKPQNLLLVKDKELLKIADLGLGRAFTVPLKSYTHEIVTLWYRAPEVLLGSTHYSTGVDMWSVGCIFAEMVRRQALFPGDSEFQQLLHIFRLLGTPTEQQWPGVSTLRDWHVYPKWEPQDLTLAVPSLSPQGVDLLTKMLKYNPAERISAKTALDHPYFDSLDKSQF 8FFCD006A9A4A75B krp2_arath Cyclin-dependent kinase inhibitor 2 KRP2 ICK2 T3A5.10 At3g50630 KIP-related protein 2 Inhibitor/interactor of CDK protein 2 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAAVRRRERDVVEENGVTTTTVKRRKMEEEVDLVESRIILSPCVQATNRGGIVARNSAGASETSVVIVRRRDSPPVEEQCQIEEEDSSVSCCSTSEEKSKRRIEFVDLEENNGDDRETETSWIYDDLNKSEESMNMDSSSVAVEDVESRRRLRKSLHETVKEAELEDFFQVAEKDLRNKLLECSMKYNFDFEKDEPLGGGRYEWVKLNP 0E5EEFE9ED256B53 krp3_arath Cyclin-dependent kinase inhibitor 3 KRP3 ICK6 K24G6.15 At5g48820 KIP-related protein 3 Inhibitor/interactor of CDK protein 6 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGKYMKKSKITGDISVMEVSKATAPSPGVRTRAAKTLALKRLNSSAADSALPNDSSCYLQLRSRRLEKPSSLIEPKQPPRVHRSGIKESGSRSRVDSVNSVPVAQSSNEDECFDNFVSVQVSCGENSLGFESRHSTRESTPCNFVEDMEIMVTPGSSTRSMCRATKEYTREQDNVIPTTSEMEEFFAYAEQQQQRLFMEKYNFDIVNDIPLSGRYEWVQVKP 7A3EB2C9A29688A7 ccd41_arath Cyclin-D4-1 At5g65420 CYCD4-1 CYCD4.1 MNA5.15 G1/S-specific cyclin-D4-1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAEENLELSLLCTESNVDDEGMIVDETPIEISIPQMGFSQSESEEIIMEMVEKEKQHLPSDDYIKRLRSGDLDLNVGRRDALNWIWKACEVHQFGPLCFCLAMNYLDRFLSVHDLPSGKGWILQLLAVACLSLAAKIEETEVPMLIDLQVGDPQFVFEAKSVQRMELLVLNKLKWRLRAITPCSYIRYFLRKMSKCDQEPSNTLISRSLQVIASTTKGIDFLEFRPSEVAAAVALSVSGELQRVHFDNSSFSPLFSLLQKERVKKIGEMIESDGSDLCSQTPNGVLEVSACCFSFKTHDSSSSYTHLS 96FD929C868DD62F krp7_arath Cyclin-dependent kinase inhibitor 7 KRP7 ICK5 F14J22.14 At1g49620 KIP-related protein 7 Inhibitor/interactor of CDK protein 5 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSETKPKRDSEYEGSNIKRMRLDDDDDVLRSPTRTLSSSSSSSLAYSVSDSGGFCSVALSEEEDDHLSSSISSGCSSSETNEIATRLPFSDLEAHEISETEISTLLTNNFRKQGISSSENLGETAEMDSATTEMRDQRKTEKKKKMEKSPTQAELDDFFSAAERYEQKRFTEKYNYDIVNDTPLEGRYQWVSLKP DIP protein Q94CL9 original sequence version: 62 6A7E0867B752E5FF krp1_arath Cyclin-dependent kinase inhibitor 1 KRP1 ICK1 F26B6.8 At2g23430 KIP-related protein 1 Inhibitor/interactor of CDK protein 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVRKYRKAKGIVEAGVSSTYMQLRSRRIVYVRSEKSSSVSVVGDNGVSSSCSGSNEYKKKELIHLEEEDKDGDTETSTYRRGTKRKLFENLREEEKEELSKSMENYSSEFESAVKESLDCCCSGRKTMEETVTAEEEEKAKLMTEMPTESEIEDFFVEAEKQLKEKFKKKYNFDFEKEKPLEGRYEWVKLE 0477A91E277C46B2 cks1_arath Cyclin-dependent kinases regulatory subunit 1 CKS1 At2g27960 T1E2.12 CKS1-At protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGQIQYSEKYFDDTFEYRHVVLPPEVAKLLPKNRLLSENEWRAIGVQQSRGWVHYAVHRPEPHIMLFRRPLNYQQQQENQAQNMLVK C8C58DEBDD7A095D krp5_arath Cyclin-dependent kinase inhibitor 5 KRP5 ICK3 K7P8.10 At3g24810 KIP-related protein 5 Inhibitor/interactor of CDK protein 3 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGKYIKKSKVAGAVSVKDKSHPPALGFRTRAAAAKNLALHRLRSHSDEADSFNYLQLRSRRLVKLPLLTNTRKQQKQQLIPSVNQCQTKNPRASSGPAKKLEPDTTTEEACGDNERISRSDCNFGDKGFDLESENRSMISDSKSIQSEIEDFFASAEQQQQRFFIQKYNFDIVSDNPLPGRYEWVKVMP A66041310E8DDED6 cdka1_arath Cyclin-dependent kinase A-1 CDKA-1 Cell division control protein 2 homolog A CDC2 At3g48750 CDC2A T21J18.20 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDQYEKVEKIGEGTYGVVYKARDKVTNETIALKKIRLEQEDEGVPSTAIREISLLKEMQHSNIVKLQDVVHSEKRLYLVFEYLDLDLKKHMDSTPDFSKDLHMIKTYLYQILRGIAYCHSHRVLHRDLKPQNLLIDRRTNSLKLADFGLARAFGIPVRTFTHEVVTLWYRAPEILLGSHHYSTPVDIWSVGCIFAEMISQKPLFPGDSEIDQLFKIFRIMGTPYEDTWRGVTSLPDYKSAFPKWKPTDLETFVPNLDPDGVDLLSKMLLMDPTKRINARAALEHEYFKDLGGMP B5FAE55FA9EC366E krp4_arath Cyclin-dependent kinase inhibitor 4 KRP4 At2g32710 ACK2 ICK7 F24L7.15 KIP-related protein 4 Inhibitor/interactor of CDK protein 7 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGKYIRKSKIDGAGAGAGGGGGGGGGGESSIALMDVVSPSSSSSLGVLTRAKSLALQQQQQRCLLQKPSSPSSLPPTSASPNPPSKQKMKKKQQQMNDCGSYLQLRSRRLQKKPPIVVIRSTKRRKQQRRNETCGRNPNPRSNLDSIRGDGSRSDSVSESVVFGKDKDLISEINKDPTFGQNFFDLEEEHTQSFNRTTRESTPCSLIRRPEIMTTPGSSTKLNICVSESNQREDSLSRSHRRRPTTPEMDEFFSGAEEEQQKQFIEKYNFDPVNEQPLPGRFEWTKVDD DIP protein Q8GYJ3 original sequence version: 60 BD7F524A308E11CF cdka_1-krp1-6 1 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association To investigate the presence and function of plant CKIs, a two-hybrid screen was performed with the Arabidopsis CDKA;1. Among the interacting clones, three putative CKIs were identified, two of which were identical to the ICK1 and ICK2 genes described previously cdka_1-krp2-2 1 13 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association To investigate the presence and function of plant CKIs, a two-hybrid screen was performed with the Arabidopsis CDKA;1. Among the interacting clones, three putative CKIs were identified, two of which were identical to the ICK1 and ICK2 genes described previously cdka_1-krp3 1 13 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association Among the positive clones, three different p27Kip1-like genes were identified: KRP1, KRP2, and KRP3. cdka_1-krp3-1 3 7 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association Table 2 cdka_1-krp1-7 3 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association Table 2 cdka_1-krp2-3 3 13 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Table 2 cdka_1-krp4 3 13 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey physical association physical association Table 2 cdka_1-krp7 3 9 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association Table 2 cdka_1-cks1 3 13 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association Table 2 krp4-cdka_1 3 14 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey physical association physical association Table 2 krp6-cdka_1-1 3 13 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association Table 2 krp7-cdka_1 3 13 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Table 2 cks1-cdka_1 3 11 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey physical association physical association Table 2 cks1-cdkb_1 3 11 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Table 2 cdkb_1-cks1 3 5 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey physical association physical association Table 2 krp4-cycd4 3 14 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Table 2 krp6-cycd4 3 4 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Table 2 krp7-cycd4 3 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Table 2 krp5-cycd4-1 3 8 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait physical association physical association Table 2 krp3-cdka_1 2 13 unspecified role unspecified role prey prey arath-cell-culture Arabidopsis thaliana cell culture plant_cell_culture plant cellular culture The culture conditions should always be cited in the experimental modifications annotation Either whole plant or undifferentiated plant cells maintained in culture. 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) physical association physical association CDKA;1 was detected in the flow-through fraction, but a significant amount also was bound to the column, indicating again a stable interaction between KRP3 and CDKA;1 Figure 2