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/ Plastid division is mediated by combinatorial assembly of plastid division proteins. Plastid division is mediated by combinatorial assembly of plastid division proteins. Plant J. (0960-7412) 2005 imex curation Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions tobac Nicotiana tabacum (Common tobacco) Common tobacco fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk Accepted 2009-OCT-1 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant J. (0960-7412) 2005 orchard Only protein-protein interactions imex curation Plastid division is mediated by combinatorial assembly of plastid division proteins. Plastid division is mediated by combinatorial assembly of plastid division proteins. Plant J. (0960-7412) 2005 imex curation Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk 27-Oct-2007: Contacted by JYOTI. orchard Only protein-protein interactions tobac Nicotiana tabacum (Common tobacco) Common tobacco bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk Accepted 2009-OCT-1 by ORCHARD. 27-Oct-2007: Contacted by JYOTI. Plant J. (0960-7412) 2005 orchard Only protein-protein interactions imex curation Plastid division is mediated by combinatorial assembly of plastid division proteins. Plastid division is mediated by combinatorial assembly of plastid division proteins. Plant J. (0960-7412) 2005 imex curation Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk 27-Oct-2007: Contacted by JYOTI. 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 Maple J., Aldridge C., Moller SG. sgm5@le.ac.uk Accepted 2009-OCT-1 by ORCHARD. Plant J. (0960-7412) 2005 orchard Only protein-protein interactions imex curation mine1_arath Cell division topological specificity factor homolog, chloroplastic MINE1 ARC12 F10D13.22 At1g69390 Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 12 F23O10.25 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAMSSGTLRISATLVSPYHHHHRNRLSLPSSSSKVDFTGFISNGVNSLETQKCTPGLAISRENTRGQVKVLARNTGDYELSPSPAEQEIESFLYNAINMGFFDRLNLAWKIIFPSHASRRSSNARIAKQRLKMILFSDRCDVSDEAKRKIVNNIIHALSDFVEIESEEKVQLNVSTDGDLGTIYSVTVPVRRVKPEYQDVDEAGTITNVEYKDTRDGSVDVRFDFYVPE FC307A9E3C4DE123 arc6_arath Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 6, chloroplastic At5g42480 ARC6 MDH9.18 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEALSHVGIGLSPFQLCRLPPATTKLRRSHNTSTTICSASKWADRLLSDFNFTSDSSSSSFATATTTATLVSPPPSIDRPERHVPIPIDFYQVLGAQTHFLTDGIRRAFEARVSKPPQFGFSDDALISRRQILQAACETLSNPRSRREYNEGLLDDEEATVITDVPWDKVPGALCVLQEGGETEIVLRVGEALLKERLPKSFKQDVVLVMALAFLDVSRDAMALDPPDFITGYEFVEEALKLLQEEGASSLAPDLRAQIDETLEEITPRYVLELLGLPLGDDYAAKRLNGLSGVRNILWSVGGGGASALVGGLTREKFMNEAFLRMTAAEQVDLFVATPSNIPAESFEVYEVALALVAQAFIGKKPHLLQDADKQFQQLQQAKVMAMEIPAMLYDTRNNWEIDFGLERGLCALLIGKVDECRMWLGLDSEDSQYRNPAIVEFVLENSNRDDNDDLPGLCKLLETWLAGVVFPRFRDTKDKKFKLGDYYDDPMVLSYLERVEVVQGSPLAAAAAMARIGAEHVKASAMQALQKVFPSRYTDRNSAEPKDVQETVFSVDPVGNNVGRDGEPGVFIAEAVRPSENFETNDYAIRAGVSESSVDETTVEMSVADMLKEASVKILAAGVAIGLISLFSQKYFLKSSSSFQRKDMVSSMESDVATIGSVRADDSEALPRMDARTAENIVSKWQKIKSLAFGPDHRIEMLPEVLDGRMLKIWTDRAAETAQLGLVYDYTLLKLSVDSVTVSADGTRALVEATLEESACLSDLVHPENNATDVRTYTTRYEVFWSKSGWKITEGSVLAS 608E776FBA73FECF mind1_arath Putative septum site-determining protein minD homolog, chloroplastic MIND1 At5g24020 ARC11 Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 11 Septum site-determining protein MinD1 MZF18.10 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASLRLFSTNHQSLLLPSSLSQKTLISSPRFVNNPSRRSPIRSVLQFNRKPELAGETPRIVVITSGKGGVGKTTTTANVGLSLARYGFSVVAIDADLGLRNLDLLLGLENRVNYTCVEVINGDCRLDQALVRDKRWSNFELLCISKPRSKLPMGFGGKALEWLVDALKTRPEGSPDFIIIDCPAGIDAGFITAITPANEAVLVTTPDITALRDADRVTGLLECDGIRDIKMIVNRVRTDMIKGEDMMSVLDVQEMLGLSLLGVIPEDSEVIRSTNRGFPLVLNKPPTLAGLAFEQAAWRLVEQDSMKAVMVEEEPKKRGFFSFFGG BE7AEF4307167825 ftz22_arath Cell division protein FtsZ homolog 2-2, chloroplastic FTSZ2-2 Plastid division protein FTSZ2-2 F3C22.150 At3g52750 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAAYVSPCLTPPDSRVLTVLRKSVLPDHHLGTRVGCLRMSEGTTKRYRVVASHKYESSSIRNSLNSHSTSHFQSQDSFLNLHPEISMLNPRKETSSVPITEDLDELSTPNTYNEARIKVIGVGGGGSNAVNRMIESEMIGVEFWIVNTDIQAMRISPVFPDNRLQIGKELTRGLGAGGNPEIGMNAATESKEAIQEALYGSDMVFVTAGMGGGTGTGGAPIIAGVAKAMGILTVGIVTTPFSFEGRRRALQAQEGIAALRDNVDTLIVIPNDKLLAAVSQSTPVTEAFNLADDILRQGVRGISDIITIPGLVNVDFADVRAIMANAGSSLMGIGTATGKTRARDAALNAIQSPLLDIGIERATGIVWNITGGSDLTLFEVNAAAEVIYDLVDPTANLIFGAVVDPSYSGQISITLIATGFKRQEEGEGRPLQATQADASMGATRRPSSSFTEGSSIEIPEFLKKKGRSRYPRL 96A32AC96CFCEDB3 ftsz1_arath Cell division protein FtsZ homolog 1, chloroplastic FTSZ1 MCO15.23 At5g55280 ARC10 FTSZ1-1 PMI4 Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 10 Protein PLASTID MOVEMENT IMPAIRED4 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAIIPLAQLNELTISSSSSSFLTKSISSHSLHSSCICASSRISQFRGGFSKRRSDSTRSKSMRLRCSFSPMESARIKVIGVGGGGNNAVNRMISSGLQSVDFYAINTDSQALLQSSAENPLQIGELLTRGLGTGGNPLLGEQAAEESKDAIANALKGSDLVFITAGMGGGTGSGAAPVVAQISKDAGYLTVGVVTYPFSFEGRKRSLQALEAIEKLQKNVDTLIVIPNDRLLDIADEQTPLQDAFLLADDVLRQGVQGISDIITIPGLVNVDFADVKAVMKDSGTAMLGVGVSSSKNRAEEAAEQATLAPLIGSSIQSATGVVYNITGGKDITLQEVNRVSQVVTSLADPSANIIFGAVVDDRYTGEIHVTIIATGFSQSFQKTLLTDPRAAKLLDKMGSSGQQENKGMSLPHQKQSPSTISTKSSSPRRLFF 82C942D597171EDB mind1 3 6 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Figure 1a demonstrating that AtMinD1 can form homodimers mind1-mine1 3 6 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey physical association physical association Figure 1a Furthermore, we found that AtMinE1 can not only form homodimers in yeast cells but that AtMinE1 also interacts with AtMinD1, suggesting the presence of an AtMinD1/AtMinE1 protein complex in Arabidopsis plastids mine1 3 4 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey physical association physical association Figure 1a Furthermore, we found that AtMinE1 can not only form homodimers in yeast cells but that AtMinE1 also interacts with AtMinD1, suggesting the presence of an AtMinD1/AtMinE1 protein complex in Arabidopsis plastids mind1-mine1-1 1 4 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role neutral component neutral component n-terminal cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 1b As in Arabidopsis (Maple et al., 2002), both AtMinE1 and AtMinD1 showed specific intraplastidic localization patterns, forming one or two discrete spots within chloroplasts mind1-2 1 6 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 1b mine1-1 1 4 unspecified role unspecified role neutral component neutral component n-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role neutral component neutral component n-terminal cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 1b mind1-mine1-2 2 4 unspecified role unspecified role neutral component neutral component c-terminal C-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal N-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 1c We divided YFP into two fragments as previously described (Hu et al., 2002b; see Experimental procedures) and fused these to the C-terminal ends of AtMinE1 or AtMinD1 to generate E-NY, E-CY, D-NY and D-CY. In cells expressing E-NY and E-CY, D-NY and E-CY or D-NY and D-CY YFP florescence indistinguishable from that observed when AtMinE1 or AtMinD1 is fused to the full length YPF was observed (Figure 1c) firmly establishing that the Min proteins can interact in planta. mine1-2 2 4 unspecified role unspecified role neutral component neutral component c-terminal N-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role neutral component neutral component c-terminal C-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range direct interaction direct interaction Figure 1c mind1-3 2 6 unspecified role unspecified role neutral component neutral component c-terminal C-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal N-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 1c mind1-mine1-3 3 4 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Figure 1a ftsz1-ftsz2 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 2a Figure 4 We found that both AtFtsZ1-1 and AtFtsZ2-1 are capable of forming homodimers, and that AtFtsZ1-1 and AtFtsZ2-1 are able to interact with each other ftsz1-ftsz2-1 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 2a ftsz1-2 3 8 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association Figure 2a Figure 4 ftsz2-2 3 7 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 2a ftsz1-ftsz2-2 1 7 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region 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 8 unspecified role unspecified role fluorescence donor fluorescence donor region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 2b The expression of F1-CFP and F2-YFP or F2-CFP and F1-YFP also yielded a positive FRET result (Figure 2b), confirming that the FtsZ proteins can interact with each other. ftsz1-ftsz2-3 1 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region 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 7 unspecified role unspecified role fluorescence donor fluorescence donor region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 2b ftsz1-4 1 8 unspecified role unspecified role fluorescence donor fluorescence donor region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region 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 direct interaction direct interaction Figure 2b ftsz2 1 7 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region 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 7 unspecified role unspecified role fluorescence donor fluorescence donor region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 2b In order to confirm that the FtsZ proteins could interact in living plant cells we utilized the FRET assay. In cells expressing F1-CFP and F1-YFP, or F2-CFP and F2-YFP a FRET signal was generated as a clear ring-like structure, demonstrating that both FtsZ proteins can form homodimers in planta ftsz2-1 2 7 unspecified role unspecified role neutral component neutral component region C-terminal region of YFP 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 7 unspecified role unspecified role neutral component neutral component region N-terminal region of YFP 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 2c o further verify the interactions between the FtsZ proteins in plant cells we utilized the BiFC assay (Hu et al., 2002b). The NY and CY fragments of YFP were fused to the C-terminal ends of AtFtsZ1-1 or AtFtsZ2-1 to generate F1-NY, F2-NY, F1-CY and F2-CY. In cells expressing both F1-NY and F1-CY or both F2-NY and F2-CY, rings of YFP fluorescence indistinguishable from those observed when the FtsZ proteins are fused to full-length YPF protein was observed (Figure 2c), demonstrating that both AtFtsZ1-1 and AtFtsZ2-1 can homodimerize in chloroplasts. Similarly, interaction between AtFtsZ1-1 and AtFtsZ2-1 was confirmed, as co-expression of F1-NY and F2-CY or F2-NY and F1-CY resulted in clear ring formation within chloroplasts ftsz1-5 2 8 unspecified role unspecified role neutral component neutral component region N-terminal region of YFP 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 8 unspecified role unspecified role neutral component neutral component region C-terminal region of YFP 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 2c ftsz1-ftsz2-4 2 7 unspecified role unspecified role neutral component neutral component region N-terminal region of YFP 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 8 unspecified role unspecified role neutral component neutral component region C-terminal region of YFP 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 2c ftsz1-ftsz2-5 2 7 unspecified role unspecified role neutral component neutral component region N-terminal region of YFP 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 8 unspecified role unspecified role neutral component neutral component region C-terminal region of YFP 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 2c ftsz2-arc6 3 5 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 7 unspecified role unspecified role bait bait physical association physical association Figure 4 further, the conserved domain of ARC6 was amplified with (amino acids 89�509) or without (amino acids 154�509) the J-domain motif, and fused to the AD to generate AD-ARC6CON and AD-ARC6ΔJ respectively. HF7c cells were co-transformed with these constructs and BD, BD-F1 or BD-F2 and interactions tested for on low-stringency media. As expected, no interactions were detected between AtFtsZ1-1 and the ARC6 domains; however, a strong interaction was detected between AtFtsZ2-1 and the conserved domain of ARC6 and this interaction was not dependent on the presence of the J-domain (Figure 4). ftsz1-6 1 8 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component c-terminal yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range direct interaction direct interaction Figure 5b To analyse the co-localization of ARC6 with AtFtsZ1-1 and AtFtsZ2-1, an ARC6-YFP fusion was co-expressed with either F1-CFP or F2-CFP. We found that transient expression of ARC6 and AtFtsZ2-1 fusions showed tight co-localization. ftsz2--arc6-1 2 5 unspecified role unspecified role neutral component neutral component region N-terminal region of YFP 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 7 unspecified role unspecified role neutral component neutral component region C-terminal region of YFP 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 5 We used BiFC assays to further analyse the specific interaction between ARC6 and AtFtsZ2-1 arc6-3 2 5 unspecified role unspecified role neutral component neutral component c-terminal region N-terminal region of YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role neutral component neutral component c-terminal C-terminal region of YFP yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 5b In most cases when ARC6-NY and ARC6-CY were transiently expressed in tobacco cells discontinuous speckled ring-like structures were observed (Figure 5b) and in other cells less organized patterns of speckles were observed (data not shown), indicating that ARC6 can interact with itself.