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/ Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. J. Integr. Plant Biol. (1672-9072) 2008 imex curation Wang F., Shi DQ., Liu J., Yang WC. wcyang@genetics.ac.cn 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. jyoti Only protein-protein interactions allce Allium cepa (Onion) Onion bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Wang F., Shi DQ., Liu J., Yang WC. Partical bombardment was used to transform onion (Allium cepa) tissue with fluorescent tagged protein expression constructs. wcyang@genetics.ac.cn Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. J. Integr. Plant Biol. (1672-9072) 2008 jyoti Only protein-protein interactions imex curation Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. J. Integr. Plant Biol. (1672-9072) 2008 imex curation Wang F., Shi DQ., Liu J., Yang WC. wcyang@genetics.ac.cn 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. 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 Wang F., Shi DQ., Liu J., Yang WC. wcyang@genetics.ac.cn Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. J. Integr. Plant Biol. (1672-9072) 2008 jyoti Only protein-protein interactions imex curation Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. Novel nuclear protein ALC-INTERACTING PROTEIN1 is expressed in vascular and mesocarp cells in Arabidopsis. J. Integr. Plant Biol. (1672-9072) 2008 imex curation Wang F., Shi DQ., Liu J., Yang WC. wcyang@genetics.ac.cn 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. 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 Wang F., Shi DQ., Liu J., Yang WC. wcyang@genetics.ac.cn Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Arabidopsis whole plant cDNA library. J. Integr. Plant Biol. (1672-9072) 2008 jyoti Only protein-protein interactions imex curation q8gwq7_arath At5g58630/mzn1_80 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MKMKSSPSPASSLSLGSKTRSFGCISIILQRFLCSGTSSTYPSDLITEPSFKSRDFIPEPLRGRGDGPGAVARLMGLDSIPVIDKTRFLSRSHSVNHLKRDDDEIQGKHRRVKSTLEYVELEDDDFFILSFEKDKNDKELGTRNCKKRRDTRGNQSKTEDQTHLEGKENNINAMNVSPGRERLEIGELMQRRKEKRKSRRKRKETKLRQNIEKENWVQSNGCWSKEEDLVNCEVRSTKEEEECGSCDDSSPVSVLDYDRYAGVRVTPSPGNISRRRLSSELESSKLNETIEQDHITLTSRSKDQEIRLHGYQEMWLMICKLTVSDLEGSDWFNGKALKLEDIEGIISEDIVSNILDQLLEETITTLSLTSQV AFF9541E5DBADF44 q9m035_arath ACI1 ALC-interacting protein 1 TON1 Recruiting Motif 29 TRM29 T10O8.80 T10O8_80 At5g01370 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTQRANPKTGCLTAVVRRLLCSGSQQTHPSDNILDSDETLQLLISYEENEEPKKEIKTETETQNDDVSLHPPPPPPPPNVVAKLMGLDHPAPGSKRFRYFDDSGSAVTRSKSVNFMDYILKGHEEEEENEDEKDGQRQCRRVKASVSFREIVPTSARWSSNNQQKKHDFLLLYLDKLDEKRELVGSSSFSRSKRFEKVVEDSKKPPLPPSAPKKKENEKVAKKFKDEPRKVAKKKKSENRNDVNGAKKVRWFLSPSKSKSSSEKTALLGESKNIRADEFIGKESESPENKSNASPVSVLDRDLYDYLILDDDYYFSGGDSESASELSTKQVETATKSSCSSSPAKTKTSTKKENNNSTNNDSEETEFITKLMNMLSDLSEEDMKSSTWVSTSSTKPVDFTQVEDFCVEFGQEILDLVMDQLVDELCI 4B89FAADBED33B0F at5g58630-at5g01370 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association Among them, 22 clones were reconfirmed in the secondary test. One of these clones, ACI1, was studied in detail because it has a nuclear localization signal (NLS) which means that it may co-localize with ALC. ACI1 (At5g01370) is predicated to encode a protein of 427 amino acids containing a lysine-rich domain, a helix-turn-helix (HTH) DNA-binding motif, a nuclear localization signal and eight proline residues in tandem at its N-terminus at5g58630-at5g01370- 3 5 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 4 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 physical association physical association To investigate which domain of ACI1 contributes to the ACI1- ALC interaction, serial truncations of ACI1 were constructed into pGBKT7 and co-transformed with the bait vector of the ALC Nterminus, respectively. Deletion experiments showed that both the lysine-rich domain and the C-terminal of ACI1 are able to interact with ALC in yeast cells, but the latter has lower affinity, indicating that ACI1 interacts with ALC via its lysine-rich domain (Figure 1C). Figure 1C alc-aci1 1 4 unspecified role unspecified role neutral component neutral component region N-terminal region of Yellow Fluorescent Protein 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 5 unspecified role unspecified role neutral component neutral component region C-terminal region of Yellow Fluorescent Protein 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 physical association physical association Figure 1D ACI1 interacts with ALC in nuclei of onion cells demonstrated by bimolecular fluorescent complementation assay.