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/ A metabolic sensor governing cell size in bacteria. A metabolic sensor governing cell size in bacteria. Cell (0092-8674) 2007 imex curation Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. plevin@biology.wustl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions 7 bacsu Bacillus subtilis fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. plevin@biology.wustl.edu Accepted 2008-FEB-8 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation A metabolic sensor governing cell size in bacteria. A metabolic sensor governing cell size in bacteria. Cell (0092-8674) 2007 imex curation Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. plevin@biology.wustl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions 7 bacsu Bacillus subtilis electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. The UgtP was labelled with Thioredoxin and 6XHis to create a Thioredoxin-UgtP-6XHis fusion protein. plevin@biology.wustl.edu Accepted 2008-FEB-8 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation A metabolic sensor governing cell size in bacteria. A metabolic sensor governing cell size in bacteria. Cell (0092-8674) 2007 imex curation Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. plevin@biology.wustl.edu 01-Feb-2008: Contacted by JYOTI. orchard Only protein-protein interactions 7 in vitro In vitro light scattering light scattering predetermined predetermined participant predetermined Weart RB., Lee AH., Chien AC., Haeusser DP., Hill NS., Levin PA. The UgtP was labelled with Thioredoxin and 6XHis to create a Thioredoxin-UgtP-6XHis fusion protein. plevin@biology.wustl.edu Accepted 2008-FEB-8 by ORCHARD. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 orchard Only protein-protein interactions 7 imex curation ezra_bacsu Septation ring formation regulator EzrA ezrA ytwP BSU29610 protein protein bacsu Bacillus subtilis MEFVIGLLIVLLALFAAGYFFRKKIYAEIDRLESWKIEILNRSIVEEMSKIKHLKMTGQTEEFFEKWREEWDEIVTAHMPKVEELLYDAEENADKYRFKKANQVLVHIDDLLTAAESSIEKILREISDLVTSEEKSREEIEQVRERYSKSRKNLLAYSHLYGELYDSLEKDLDEIWSGIKQFEEETEGGNYITARKVLLEQDRNLERLQSYIDDVPKLLADCKQTVPGQIAKLKDGYGEMKEKGYKLEHIQLDKELENLSNQLKRAEHVLMTELDIDEASAILQLIDENIQSVYQQLEGEVEAGQSVLSKMPELIIAYDKLKEEKEHTKAETELVKESYRLTAGELGKQQAFEKRLDEIGKLLSSVKDKLDAEHVAYSLLVEEVASIEKQIEEVKKEHAEYRENLQALRKEELQARETLSNLKKTISETARLLKTSNIPGIPSHIQEMLENAHHHIQETVNQLNELPLNMEEAGAHLKQAEDIVNRASRESEELVEQVILIEKIIQFGNRFRSQNHILSEQLKEAERRFYAFDYDDSYEIAAAAVEKAAPGAVEKIKADISA 9AD2D58A76A08FDF ftsz_bacsu Cell division protein FtsZ ftsZ BSU15290 protein protein bacsu Bacillus subtilis MLEFETNIDGLASIKVIGVGGGGNNAVNRMIENEVQGVEYIAVNTDAQALNLSKAEVKMQIGAKLTRGLGAGANPEVGKKAAEESKEQIEEALKGADMVFVTAGMGGGTGTGAAPVIAQIAKDLGALTVGVVTRPFTFEGRKRQLQAAGGISAMKEAVDTLIVIPNDRILEIVDKNTPMLEAFREADNVLRQGVQGISDLIATPGLINLDFADVKTIMSNKGSALMGIGIATGENRAAEAAKKAISSPLLEAAIDGAQGVLMNITGGTNLSLYEVQEAADIVASASDQDVNMIFGSVINENLKDEIVVTVIATGFIEQEKDVTKPQRPSLNQSIKTHNQSVPKREPKREEPQQQNTVSRHTSQPADDTLDIPTFLRNRNKRG 24E908DEC9685614 ugtp_bacsu Processive diacylglycerol beta-glucosyltransferase UDP glucosyltransferase UDP-glucose:1,2-diacylglycerol-3-beta-D-glucosyltransferase Beta-gentiobiosyldiacylglycerol synthase ugtP ypfP BSU21920 Beta-diglucosyldiacylglycerol synthase Beta-monoglucosyldiacylglycerol synthase Beta-triglucosyldiacylglycerol synthase Glucosyl-beta-1,6-glucosyldiacylglycerol synthase Diglucosyl diacylglycerol synthase (1,6-linking) protein protein bacsu Bacillus subtilis MNTNKRVLILTANYGNGHVQVAKTLYEQCVRLGFQHVTVSNLYQESNPIVSEVTQYLYLKSFSIGKQFYRLFYYGVDKIYNKRKFNIYFKMGNKRLGELVDEHQPDIIINTFPMIVVPEYRRRTGRVIPTFNVMTDFCLHKIWVHENVDKYYVATDYVKEKLLEIGTHPSNVKITGIPIRPQFEESMPVGPIYKKYNLSPNKKVLLIMAGAHGVLKNVKELCENLVKDDQVQVVVVCGKNTALKESLSALEAENGDKLKVLGYVERIDELFRITDCMITKPGGITLTEATAIGVPVILYKPVPGQEKENANFFEDRGAAIVVNRHEEILESVTSLLADEDTLHRMKKNIKDLHLANSSEVILEDILKESEMMTAKQKAKVLS 8A8B98593DAC6041 ugtp-ezra 1 6 unspecified role unspecified role neutral component neutral component 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 bacsu Bacillus subtilis 4 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 bacsu Bacillus subtilis colocalization colocalization 4D Colocalization of YFP-UgtP and a CFP fusion to the cell division protein EzrA (which functions as a proxy for FtsZ localization [Levin et al., 1999; Wu and Errington, 2004]) confirmed that 84% (42/50) of UgtP foci colocalize with FtsZ at midcell in wild-type cells. In contrast, the majority of UgtP puncta in pgcA::Tn10 cells (67.2%, 45/ 67) are not associated with FtsZ (Figure 4D). ftsza-ftsza 3 5 unspecified role unspecified role neutral component neutral component in vitro In vitro 6 inhibitor inhibitor neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined thioredoxin fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Figure 3 A-D Thio-Ugtp was able to directly inhibit FtsZ assembly in-vitro at ratios as low as 1:5. To test whether UgtP inhibits FtsZ assembly directly, we examined the effect of UgtP on FtsZ assembly in vitro. For these experiments we employed native B. subtilis FtsZ and a Thioredoxin-UgtP-6XHis fusion protein (Thio-UgtP). FtsZ assembly was measured using a 90 degree angle light-scattering assay (Mukherjee and Lutkenhaus, 1999). Using this approach we determined that UgtP inhibited FtsZ assembly in a concentration-dependent manner with 90% inhibition at a 1:1 ratio of Thio-UgtP to FtsZ (Figures 3A and 3B). Thio-UgtP was able to inhibit FtsZ assembly at ratios as low as 1:5 (Figure 3B). The absence of UgtP's substrate, UDP-Glc, resulted in a small but statistically significant reduction in its ability to inhibit FtsZ assembly in vitro (Figure 3C). Adding 6 mM Thioredoxin-6XHis (Thio) or BSA had no effect on FtsZ assembly (Figure 3D). The participant EBI-1569853(P17865) was duplicated. It is posssible that originally this interaction did contain more participants pointing to different Trembl proteins which have been merged to a single swissprot protein later. ftsza-ftsza-1 3 5 unspecified role unspecified role neutral component neutral component in vitro In vitro 5 unspecified role unspecified role neutral component neutral component in vitro In vitro physical association physical association Figure 3 A-D FtsZ protofilaments were shown to assemble in-vitro. FtsZ assembly was measured using a 90 degree angle light-scattering assay (Mukherjee and Lutkenhaus, 1999). ftsza-ftsza-2 2 5 unspecified role unspecified role neutral component neutral component in vitro In vitro 5 unspecified role unspecified role neutral component neutral component in vitro In vitro physical association physical association Figure 3 E FtsZ protofilaments were shown to assemble in-vitro. Electron micrographs of FtsZ assembled in the presence and absence of UgtP suggest that UgtP interferes with stabilizing lateral interactions between single-stranded protofilaments. In the absence of UgtP, FtsZ formed large bundled structures, the consequence of lateral interactions between protofilaments (Figure 3E). ftsza-ftsza-3 2 6 inhibitor inhibitor neutral component neutral component in vitro In vitro 5 unspecified role unspecified role neutral component neutral component in vitro In vitro physical association physical association Figure 3 F Thio-Ugtp was able to inhibit FtsZ assembly in-vitro at a ratio of 1:2, as only short polymers were present with little bundling observed. However, we observed very few bundles at a 1:2 ratio of UgtP to FtsZ, and protofilaments were significantly shorter, consistent with a decrease in polymer stability (Figure 3F). The participant EBI-1569853(P17865) was duplicated. It is posssible that originally this interaction did contain more participants pointing to different Trembl proteins which have been merged to a single swissprot protein later. ugtp-ugtp 2 6 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined thioredoxin fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined thioredoxin fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Figure 3G, 3H UgtP itself was able to form higher order structures in FtsZ assembly buffer (Figures 3G and 3H). Intriguingly, UgtP itself formed higher order structures in FtsZ assembly buffer (Figures 3G and 3H). These structures, which appear to consist of closely stacked spirals of UgtP monomers (seven or eight per turn), were less prevalent in the presence of FtsZ (Figure 3F), suggesting that UgtP inhibition of FtsZ assembly occurs at the expense of UgtP self-assembly. ugtp-ugtp-1 2 5 inhibitor inhibitor neutral component neutral component in vitro In vitro 6 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined thioredoxin fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Figure 3F, 3G UgtP's ablility to to inhibit FtsZ assembly in-vitro occurs at the expence of UgtP self-assembly. Intriguingly, UgtP itself formed higher order structures in FtsZ assembly buffer (Figures 3G and 3H). These structures, which appear to consist of closely stacked spirals of UgtP monomers (seven or eight per turn), were less prevalent in the presence of FtsZ (Figure 3F), suggesting that UgtP inhibition of FtsZ assembly occurs at the expense of UgtP self-assembly. The participant EBI-1567571(P54166) was duplicated. It is posssible that originally this interaction did contain more participants pointing to different Trembl proteins which have been merged to a single swissprot protein later.