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/ The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. J. Biol. Chem. (0021-9258) 2008 imex curation Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr 16-Oct-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 Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr Accepted 2008-SEP-1 by CDEROW. 16-Oct-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2008 orchard Only protein-protein interactions imex curation The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. J. Biol. Chem. (0021-9258) 2008 imex curation Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr 16-Oct-2008: Contacted by JYOTI. Only protein-protein interactions 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. bn-page blue native page bn-page western blot western blot Immuno blot Immuno blot Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr Accepted 2008-SEP-1 by CDEROW. 16-Oct-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. The three mitochondrial encoded CcmF proteinsform a complex that interacts with CcmH and C-type apo-cytochromes in Arabidopsis. J. Biol. Chem. (0021-9258) 2008 imex curation Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr 16-Oct-2008: Contacted by JYOTI. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast ub reconstruction ubiquitin reconstruction ub reconstruction predetermined predetermined participant predetermined Rayapuram N., Hagenmuller J., Grienenberger JM., Bonnard G., Giege P. Phillippe.Giege@ibmp-ulp.u-strasbg.fr Accepted 2008-SEP-1 by CDEROW. 16-Oct-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation cyc1_arath Cytochrome c-1 Cytochrome c At1g22840 F19G10.20 At1g22840 CYTC-1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASFDEAPPGNAKAGEKIFRTKCAQCHTVEAGAGHKQGPNLNGLFGRQSGTTAGYSYSAANKNKAVEWEEKALYDYLLNPKKYIPGTKMVFPGLKKPQDRADLIAYLKESTAPK AB9E7BD40AFF2C62 ccmh_arath Cytochrome c-type biogenesis CcmH-like mitochondrial protein CCMH F9L1.17 At1g15220 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEKTDEERKKAQMLDARARNISHNVRCTECGSQSIEDSQADIAILLRQLIRNEIGAGKTDKEIYSKLEDEFGETVLYAPKFDLQTAALWLTPVIIAGGTAAGIVYQKHRLRKNVDIMALNLIRGVPLTPKERVTILDVLIPPSPPPQGVVSRLRRWLNR 51637FB6AFD683F7 ccmfc_arath Cytochrome c biogenesis CcmF C-terminal-like mitochondrial protein CCMFC CCB452 CCB6C Cytochrome c biogenesis orf452 AtMg00180 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVQLHNFFFFIIFMVVLCGTAAPVLLKWFVSRDVSTGAPFFNGTIIPILISLFSLLVYLHSRKIIRSMDGAKSGVLVRASRPILLPDIIGRSSSETRARKALFFFVPVLHFCLLESKGDFSYLESFCGVLCLLFFCTFFFLARDRSAKRERARRRKGQTLRPNGNEQRRNDKMRCSGHPHLDLERRVEGFGPLAFPVPPELGGACVGGVPPEIGLEALALPRSRQLMAMAVGHDYYQKVPMKMNISHGGVCICMLGVLLSNTKKIQFTQRLPLGYELHMGKERCCLRGLDHLHGPTFHSICGNLMIYKSSLTNDRLMFEHDESLHADLLLINFPASYKNGKLEHFLHWWMKNRKHNNFWLTMFPEKRYFRERTSTAEVAIHTNLFTDLYALIGTGSSRTGGWYTTIMKLPFIFFIWIGFMLASLGGLPSLLRQLQKDKLRWN AFDAA24F80423BD4 ccmf1_arath Cytochrome c biogenesis CcmF N-terminal-like mitochondrial protein 1 CCMFN1 CC6BN1 CCB382 Cytochrome c biogenesis orf382 AtMg00830 At2g07783 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSISIYEFFHYSLFLGLFVAFTYNKKQPPAFGAALAFWCILLSFLGLLFCHISNNLSNYNVLTANALFFYQISGTWSNHEGSILLWCWILNFYGFFFCYRGRPQSHNVLKQGGYRESLFFFFVLNFVKNSILSLLRYEQESGLKNQLYTPFVLRTLVDSELCSRRNRTFDGPALFYAPLYPERKIKNPLDAWRFRGSREGKRTHPLLHLARDDKERASSIDEQRIDGALGIALFFSFFLLASSDPFVRNFFVCTEPLAELNPVLQDPILAIHPPCIYAGDVASAEGFGLCRSKMMNGIVALHSPPMRKDAAEKNGTLLCSAGCVGSRITSELFTLKFKHVGAKCYPALLLRSNRSPLMLLRRRFFAFSSFWAGARSHSTKRY 7B9100599199E244 ccmf2_arath Cytochrome c biogenesis CcmF N-terminal-like mitochondrial protein 2 CCMFN2 CC6BN2 CCB203 Cytochrome c biogenesis orf203 AtMg00960 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress VDTGREQAKRVVRNGKKETTTLPLCWTAGANTVVSDQDQEPIRIWILTCWWFLTVGILLGSWWAYHELGWGGWWFWDPVENASFMPWVLATACIHSVILPFLHSWTLFLNIVTFLCCVLGTFSIRSGLLASVHSFATDDTRGIFLWWFFLLMTGIFMILFYQMKQQASVRRTYKKEMVVARSTLVHLRHSARAQPRPVMLWKN AF3282E8F7524B81 cyc1b_arath Cytochrome c1 2, heme protein, mitochondrial CYC1-2 Complex III subunit 4-2 Complex III subunit IV-2 Cytochrome b-c1 complex subunit 4-2 Ubiquinol-cytochrome c reductase complex cytochrome c1 subunit 2 MHK7.4 At5g40810 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVGGGVIRQLLRRKLHSQSVATPVLSWLSSKKANEDAGSAGLRAFALMGAGITGLLSFSTVASADEAEHGLECPNYPWPHEGILSSYDHASIRRGHQVYQQVCASCHSMSLISYRDLVGVAYTEEEAKAMAAEIEVVDGPNDEGEMFTRPGKLSDRLPEPYSNESAARFANGGAYPPDLSLVTKARHNGQNYVFALLTGYRDPPAGISIREGLHYNPYFPGGAIAMPKMLNDEAVEYEDGTPATEAQMGKDVVSFLSWAAEPEMEERKLMGFKWIFLLSLALLQAAYYRRLKWSVLKSRKLVLDVVN 3F807D081F788ED3 ccmfn1-ccmfn2 2 7 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Figure 3 This does not prove a direct interaction between the 3 Arabidopsis CcmF proteins but strongly suggests that they are, together with other proteins, in particular CCMH that was also found in a 500 kDa complex The complex was judged to be approximately 500kDa in size. ccmfn1-ccmfn2-1 3 8 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A For FN1 Cub, we observed growth with FN2 constructs, ccmfn1-ccmfn2-2 3 8 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A For FN1 Cub, we observed growth with FN2 constructs, ccmfn2-ccmfn1 3 8 unspecified role unspecified role bait bait n-terminal CubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A for FN2 Cub, growth was observed with both FN1 and FC constructs The authors concluded "CcmFN2 can interact with both CcmFN1 and CcmFC and thus suggest that the 3 mitochondrial encoded Arabidopsis CcmF proteins are assembled in vivo into a protein complex that reconstitute a bacterial-like CcmF". ccmfn2-ccmfn1-1 3 7 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 unspecified role unspecified role bait bait n-terminal CubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 4A for FN2 Cub, growth was observed with both FN1 and FC constructs The authors concluded "CcmFN2 can interact with both CcmFN1 and CcmFC and thus suggest that the 3 mitochondrial encoded Arabidopsis CcmF proteins are assembled in vivo into a protein complex that reconstitute a bacterial-like CcmF". ccmfn2-ccmfc-1 3 8 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 4A This shows that CcmFN2 can interact with CcmFN1 and CcmFC whereas CcmFN1 cannot interact with CcmFC. The authors concluded "CcmFN2 can interact with both CcmFN1 and CcmFC and thus suggest that the 3 mitochondrial encoded Arabidopsis CcmF proteins are assembled in vivo into a protein complex that reconstitute a bacterial-like CcmF". ccmfn2-ccmfc-2 3 8 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A This shows that CcmFN2 can interact with CcmFN1 and CcmFC whereas CcmFN1 cannot interact with CcmFC. The authors concluded "CcmFN2 can interact with both CcmFN1 and CcmFC and thus suggest that the 3 mitochondrial encoded Arabidopsis CcmF proteins are assembled in vivo into a protein complex that reconstitute a bacterial-like CcmF". ccmfn2-at1g22840 3 4 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. The authors concluded "CcmFN2 can interact with both c-type apo-cytochromes and suggest that in vivo the assembled CcmF indeed interacts with ctype apo-cytocromes at the level of CcmFN2.". ccmfn2-at1g22840-1 3 8 unspecified role unspecified role bait bait n-terminal CubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. The authors concluded "CcmFN2 can interact with both c-type apo-cytochromes and suggest that in vivo the assembled CcmF indeed interacts with ctype apo-cytocromes at the level of CcmFN2.". ccmfn2-at5g40810 3 8 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. The authors concluded "CcmFN2 can interact with both c-type apo-cytochromes and suggest that in vivo the assembled CcmF indeed interacts with ctype apo-cytocromes at the level of CcmFN2.". ccmfn2-at5g40810-1 3 9 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role bait bait n-terminal CubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. The authors concluded "CcmFN2 can interact with both c-type apo-cytochromes and suggest that in vivo the assembled CcmF indeed interacts with ctype apo-cytocromes at the level of CcmFN2.". at1g15220-ccmfn1-2 3 7 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 6A First, split ubiquitin assays showed that CCMH Cub fusion proteins could interact with both CcmFN1 and CcmFN2 Nub fusions, but not with the CcmFC fusion at1g15220-ccmfn2-3 3 5 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 6A First, split ubiquitin assays showed that CCMH Cub fusion proteins could interact with both CcmFN1 and CcmFN2 Nub fusions, but not with the CcmFC fusion ccmfc-ccmfn2 3 8 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A This shows that CcmFN2 can interact with CcmFN1 and CcmFC whereas CcmFN1 cannot interact with CcmFC. ccmfc-ccmfn2-1 3 6 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 4A This shows that CcmFN2 can interact with CcmFN1 and CcmFC whereas CcmFN1 cannot interact with CcmFC. at5g40810-ccmfn2 3 8 unspecified role unspecified role prey prey n-terminal NubPLV tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. at5g40810-ccmfn2-1 3 8 unspecified role unspecified role prey prey c-terminal NubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role bait bait c-terminal CubPLV tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 5 FN2 Cub growth was observed with both apocytocrome c and apocytochrome c1 constructs. ccmfn1-at1g15220 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 7 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 6B The activation of reporter genes was observed whenthe CCMH-D1 AD construct was co-transformed with CcmFN1 domain 5 and with CcmFN2 domain 2 BD constructs but not with CcmFN1 domain 3 and CcmFC domain 6 BD constructs The range for CCMH D1 domain was taken from Meyer et al Proc Natl Acad Sci U S A. 2005 November 1; 102(44): 16113-16118. PMID 16236729. The authors concluded "CCMH can interact with CcmF at the level of both CcmFN1 and CcmFN2 and that these interactions are, at least in part, mediated by two domains localized in the inter-membrane space in CcmFN1 and CcmFN2". ccmfn2-at1g15220 1 8 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 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 6B The activation of reporter genes was observed when the CCMH-D1 AD construct was co-transformed with CcmFN1 domain 5 and with CcmFN2 domain 2 BD constructs but not with CcmFN1 domain 3 and CcmFC domain 6 BD constructs The authors concluded "CCMH can interact with CcmF at the level of both CcmFN1 and CcmFN2 and that these interactions are, at least in part, mediated by two domains localized in the inter-membrane space in CcmFN1 and CcmFN2". The range for CCMH D1 domain was taken from Meyer et al Proc Natl Acad Sci U S A. 2005 November 1; 102(44): 16113-16118. PMID 16236729.