MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. FEBS Lett. (0014-5793) 2013 Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. bacsu Bacillus subtilis anti bait coip anti bait coimmunoprecipitation anti bait coip experimental particp experimental participant identification experimental particp Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. FEBS Lett. (0014-5793) 2013 Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. FEBS Lett. (0014-5793) 2013 Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. FEBS Lett. (0014-5793) 2013 Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. Transcription factor TnrA inhibits the biosynthetic activity of glutamine synthetase in Bacillus subtilis. FEBS Lett. (0014-5793) 2013 Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Fedorova K., Kayumov A., Woyda K., Ilinskaja O., Forchhammer K. FEBS Lett. (0014-5793) 2013 tnra_bacsu HTH-type transcriptional regulator TnrA tnrA scgR BSU13310 Transcriptional nitrogen regulatory protein A protein protein bacsu Bacillus subtilis MTTEDHSYKDKKVISIGIVSELTGLSVRQIRYYEERKLIYPQRSSRGTRKYSFADVERLMDIANKREDGVQTAEILKDMRKKEQMLKNDPQVRKKMLEGQLNAHFRYKNR mint Regulator of transcription that activates or represses many genes during nitrogen-limited growth; among others, it activates nrgAB, nasABCDE, gabP, and represses glnRA 201AA41E76E0E631 glnk_bacsu Sensor histidine kinase GlnK glnK ycbA yzgA BSU02440 nrgB protein protein bacsu Bacillus subtilis MLITVPLAGELKFYPLNEEFRVSFGAPVFFFFLSLLRHVPAVLPGFLTGAAVFIFRVFLELWGGGHNGLTPILYDQASGFFFYMTYACLFSILKANRFRERPIMLGFIGFMIEVVSDCVELTVQFLIFHTVVTPEKITDIAVIAISHTFIVMSFYSVLKLYETQSREKQTRQQHEHMLMIVSNLYEETVHLKKTLKTTEKVTNDSYQLYREMKGKDVQLSGRILRLAGEIHEVKKDNQRIFAGLSKLISNESLRDYMRASDLLQLVIRMNEKYAEALGKQIDFYCSIEGEHDEYHVFIVLSIINNLTANAVEAMDEEGMVSLRLRKPNESMVEFQVEDNGPGISEKIGDIVFDPGFTSKYDEFGTPSTGIGLSYVKEIVTELEGDITFDNQQRGVVFAIRLPVRHLIQKG mint 88D0E033366113E2 gln1a_bacsu Glutamine synthetase Glutamine synthetase I alpha BSU17460 Glutamate--ammonia ligase glnA protein protein bacsu Bacillus subtilis MAKYTREDIEKLVKEENVKYIRLQFTDILGTIKNVEIPVSQLGKALDNKVMFDGSSIEGFVRIEESDMYLYPDLNTFVIFPWTAEKGKVARFICDIYNPDGTPFEGDPRNNLKRILKEMEDLGFSDFNLGPEPEFFLFKLDEKGEPTLELNDKGGYFDLAPTDLGENCRRDIVLELEEMGFEIEASHHEVAPGQHEIDFKYAGAVRSCDDIQTFKLVVKTIARKHGLHATFMPKPLFGVNGSGMHCNLSLFKNGVNAFFDENADLQLSETAKHFIAGIVKHATSFTAVTNPTVNSYKRLVPGYEAPCYVAWSAQNRSPLIRIPASRGISTRVEVRSVDPAANPYLALSVLLAAGLDGIKNKLEAPAPIDRNIYVMSKEERMENGIVDLPATLAEALEEFKSNEVMVKALGEHLFEHFIEAKEIEWDMFRTQVHPWEREQYMSQY 83A5657CE1388AB0 83A5657CE1388AB0 tnra-glna 3 TnrA 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain his tag 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 GS 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified direct interaction direct interaction mint f6 sf3 To prove competitive binding of GlnK and GS to TnrA in vitro binding was analyzed by SPR spectroscopy [23]. Full length His6- TnrA was immobilized on a sensor chip and His6- TnrA35, which is not able to interact with GS and GlnK was used as a control. Since the SPR signal is proportional to the mass of the ligand, binding of the large GS protein (600 kDa) to TnrA results in a much larger increase in resonance units than binding of GlnK protein (36 kDa). Injecting a mixture of GS and GlnK to the TnrA-loaded sensor-chip should reveal the competition of GlnK and GS for TnrA binding due to lowering the SPR signal. glna-tnra 1 TnrA 4 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level GS 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f5 mint To examine the correlation between the measured in situ GS activities and the interactions of TnrA with either GlnK or GS before and after glutamine addition, immunoprecipitation experiments with GS or GlnK-specific antibodies were performed as described in supplementary materials, and the presence of TnrA in the immunoprecipitate was evaluated by TnrA-specific western-blotting (Fig. 5). In wild type nitrate grown cells, TnrA was completely associated with GlnK before glutamine addition, while after glutamine addition it was only associated with GS, in agreement with earlier data [17, 27]. In the AmtB-deficient strain, the same change of TnrA localization from GlnK to GS was observed. This indicates that the AmtB protein does not participate in the control of TnrA interactions. TnrA, which is bound to cytoplasmic GlnK before glutamine addition, is able to swap to GS after glutamine addition, which correlates with the depression of in situ GS activity. In the GlnK-deficient strain, TnrA was bound to GS before and after glutamine addition, which correlates with the constitutive GS activity during the glutamine shift experiments. glnk-tnra 1 TnrA 4 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level GlnK 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f5 mint To examine the correlation between the measured in situ GS activities and the interactions of TnrA with either GlnK or GS before and after glutamine addition, immunoprecipitation experiments with GS or GlnK-specific antibodies were performed as described in supplementary materials, and the presence of TnrA in the immunoprecipitate was evaluated by TnrA-specific western-blotting (Fig. 5). In wild type nitrate grown cells, TnrA was completely associated with GlnK before glutamine addition, while after glutamine addition it was only associated with GS, in agreement with earlier data [17, 27]. In the AmtB-deficient strain, the same change of TnrA localization from GlnK to GS was observed. This indicates that the AmtB protein does not participate in the control of TnrA interactions. TnrA, which is bound to cytoplasmic GlnK before glutamine addition, is able to swap to GS after glutamine addition, which correlates with the depression of in situ GS activity. In the GlnK-deficient strain, TnrA was bound to GS before and after glutamine addition, which correlates with the constitutive GS activity during the glutamine shift experiments. tnra-glna-1 2 TnrA 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified his tag 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain GS 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction sf1 mint glnk-glna 2 GlnK 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified his tag 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain GS 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction sf1 mint