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 DNA replication stress differentially regulates G1/S genes via Rad53-dependent inactivation of Nrm1. DNA replication stress differentially regulates G1/S genes via Rad53-dependent inactivation of Nrm1. EMBO J. (0261-4189) 2012 imex curation Travesa A., Kuo D., de Bruin RA., Kalashnikova TI., Guaderrama M., Thai K., Aslanian A., Smolka MB., Yates JR., Ideker T., Wittenberg C. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Travesa A., Kuo D., de Bruin RA., Kalashnikova TI., Guaderrama M., Thai K., Aslanian A., Smolka MB., Yates JR., Ideker T., Wittenberg C. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation rad53_yeast Serine/threonine-protein kinase RAD53 RAD53 MEC2 SAD1 Serine-protein kinase 1 SPK1 P2588 YPL153C CHEK2 homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MENITQPTQQSTQATQRFLIEKFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSIKKVWTFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSNQLLSQGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDRIRSNLKNTSKIASPGLTSSTASSMVANKTGIFKDFSIIDEVVGQGAFATVKKAIERTTGKTFAVKIISKRKVIGNMDGVTRELEVLQKLNHPRIVRLKGFYEDTESYYMVMEFVSGGDLMDFVAAHGAVGEDAGREISRQILTAIKYIHSMGISHRDLKPDNILIEQDDPVLVKITDFGLAKVQGNGSFMKTFCGTLAYVAPEVIRGKDTSVSPDEYEERNEYSSLVDMWSMGCLVYVILTGHLPFSGSTQDQLYKQIGRGSYHEGPLKDFRISEEARDFIDSLLQVDPNNRSTAAKALNHPWIKMSPLGSQSYGDFSQISLSQSLSQQKLLENMDDAQYEFVKAQRKLQMEQQLQEQDQEDQDGKIQGFKIPAHAPIRYTQPKSIEAETREQKLLHSNNTENVKSSKKKGNGRFLTLKPLPDSIIQESLEIQQGVNPFFIGRSEDCNCKIEDNRLSRVHCFIFKKRHAVGKSMYESPAQGLDDIWYCHTGTNVSYLNNNRMIQGTKFLLQDGDEIKIIWDKNNKFVIGFKVEINDTTGLFNEGLGMLQEQRVVLKQTAEEKDLVKKLTQMMAAQRANQPSASSSSMSAKKPPVSDTNNNGNNSVLNDLVESPINANTGNILKRIHSVSLSQSQIDPSKKVKRAKLDQTSKGPENLQFS 84A9612229CA72D1 84A9612229CA72D1 atg18_yeast Autophagy-related protein 18 NMR1 YFR021W ATG18 AUT10 CVT18 SVP1 Cytoplasm to vacuole targeting protein 18 Swollen vacuole phenotype protein 1 Needed for premeiotic replication protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSDSSPTINFINFNQTGTCISLGTSKGFKIFNCEPFGKFYSEDSGGYAIVEMLFSTSLLALVGIGDQPALSPRRLRIINTKKHSIICEVTFPTSILSVKMNKSRLVVLLQEQIYIYDINTMRLLHTIETNPNPRGLMAMSPSVANSYLVYPSPPKVINSEIKAHATTNNITLSVGGNTETSFKRDQQDAGHSDISDLDQYSSFTKRDDADPTSSNGGNSSIIKNGDVIVFNLETLQPTMVIEAHKGEIAAMAISFDGTLMATASDKGTIIRVFDIETGDKIYQFRRGTYATRIYSISFSEDSQYLAVTGSSKTVHIFKLGHSMSNNKLDSDDSNMEEAAADDSSLDTTSIDALSDEENPTRLAREPYVDASRKTMGRMIRYSSQKLSRRAARTLGQIFPIKVTSLLESSRHFASLKLPVETNSHVMTISSIGSPIDIDTSEYPELFETGNSASTESYHEPVMKMVPIRVVSSDGYLYNFVMDPERGGDCLILSQYSILMD 06B2DFAF842AE933 06B2DFAF842AE933 atg18-rad53 1 RAD53 2 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined NMR1 3 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra predetermined predetermined participant predetermined enzyme target enzyme target substrate substrate neutral component neutral component purified purified phosphorylated phosphorylated residue phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain resulting-ptm phosphorylated phosphorylated residue phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain resulting-ptm undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:38 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' Ser47Ala undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:41 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' Thr42Ala undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:41 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' Ser240Ala phosphorylated phosphorylated residue phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain resulting-ptm undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:33:45 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' Ser145Ala gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phosphorylation phosphorylation reaction phosphorylation homomint mint To examine that possibility, we evaluated the capacity of bacterially expressed, enzymatically active Rad53 to phosphorylate GST Nrm1 purified from bacteria (Figure 5C). The activity of Rad53 was assessed as the capacity of Rad53 to autophosphorylate itself (top panel, Figure 5C). Rad53 phosphorylated GST Nrm1, but not GST alone, in vitro in the absence of other yeast proteins. In contrast, an enzymatically inactive mutant of Rad53 fails to phosphorylate GST Nrm1. Next, residues that were directly phosphorylated by Rad53 were identified by mass spectrometry using GST Nrm1 phosphorylated in vitro in a Rad53 kinase assay performed with unlabelled ATP, as described above (see Materials and methods). That approach identified three phosphorylated residues with high confidence, T42, S47 and S240. f5c f6d f6e