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 A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions in vitro In vitro in gel kinase assay in-gel kinase assay in gel kinase assay predetermined predetermined participant predetermined Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. 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 Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. 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 Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. I have annotated the features of amino acid substitutions as 'experimental feature' because the wt protein was not used as a comparison in the screens of the cDNA libraries. It was therefore not demonstrated that the mutant forms of the protein were more or less effective in binding the prey. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. A novel yeast two-hybrid approach to identify CDPK substrates: characterization of the interaction between AtCPK11 and AtDi19, a nuclear zinc finger protein. FEBS Lett. (0014-5793) 2006 imex curation Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. Only protein-protein interactions arath-leaf Arabidopsis thaliana leaf leaf leaf fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Rodriguez Milla MA., Uno Y., Chang IF., Townsend J., Maher EA., Quilici D., Cushman JC. Both catalytically impaired and constitutively active forms of CPK11 protein were used as baits in the yeast two hybrid assay. jcushman@unr.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI Arabidopsis three day old etiolated seedling cDNA libraray. A pooled cDNA library of arabidopsis three week old in vitro grown seedlings subjected to one of the following treatments 1.100 microM abscissic acid, 2. 100 microM gibberillic acid, 3. 100 microM 2,4-D, 4. cold treatment (4C), 5. 100 microM BAP, 6. anaerobiosis, 7. salicylic acid, 8. dessication, 9. heat (42C), 10. 24h dark, 11. wounding, 12. high light, 13. 150 mM NaCl, 14. 100 microM IAA, 15. 100 microM methyl jasmonate, 16. 440 mM mannitol. FEBS Lett. (0014-5793) 2006 Only protein-protein interactions imex curation cdpkc_arath Calcium-dependent protein kinase 12 CPK12 At5g23580 CDPK9 MQM1.15 Calcium-dependent protein kinase isoform CDPK9 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MANKPRTRWVLPYKTKNVEDNYFLGQVLGQGQFGTTFLCTHKQTGQKLACKSIPKRKLLCQEDYDDVLREIQIMHHLSEYPNVVRIESAYEDTKNVHLVMELCEGGELFDRIVKRGHYSEREAAKLIKTIVGVVEACHSLGVVHRDLKPENFLFSSSDEDASLKSTDFGLSVFCTPGEAFSELVGSAYYVAPEVLHKHYGPECDVWSAGVILYILLCGFPPFWAESEIGIFRKILQGKLEFEINPWPSISESAKDLIKKMLESNPKKRLTAHQVLCHPWIVDDKVAPDKPLDCAVVSRLKKFSAMNKLKKMALRVIAERLSEEEIGGLKELFKMIDTDKSGTITFEELKDSMRRVGSELMESEIQELLRAADVDESGTIDYGEFLAATIHLNKLEREENLVAAFSFFDKDASGYITIEELQQAWKEFGINDSNLDEMIKDIDQDNDGQIDYGEFVAMMRKGNGTGGGIGRRTMRNSLNFGTTLPDESMNV 0315346396585264 di191_arath Protein DEHYDRATION-INDUCED 19 DI19-1 At1g56280 DI19 F14G9.11 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDADSKRFLATLRSRSEMLMGFEEIDGDDDFQEEFACPFCAESYDIIGLCCHIDDEHTLESKNAVCPVCSLKVGVDIVAHITLHHGSLFKLQRKRKSRKSGTNSTLSLLRKELREGDLQRLLGFTSRNGSVASSVTPDPLLSSFISPTRSQSSPAPRQTKNVSEDKQIERKRQVFISPVSLKDREERRHKSEFVQRLLSSAIFDEV DIP protein Q39083 original sequence version: 74 F1E96902AB60B74B cdpk4_arath Calcium-dependent protein kinase 4 CPK4 T25P22.10 At4g09570 Calmodulin-domain protein kinase CDPK isoform 4 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEKPNPRRPSNSVLPYETPRLRDHYLLGKKLGQGQFGTTYLCTEKSSSANYACKSIPKRKLVCREDYEDVWREIQIMHHLSEHPNVVRIKGTYEDSVFVHIVMEVCEGGELFDRIVSKGCFSEREAAKLIKTILGVVEACHSLGVMHRDLKPENFLFDSPSDDAKLKATDFGLSVFYKPGQYLYDVVGSPYYVAPEVLKKCYGPEIDVWSAGVILYILLSGVPPFWAETESGIFRQILQGKIDFKSDPWPTISEGAKDLIYKMLDRSPKKRISAHEALCHPWIVDEHAAPDKPLDPAVLSRLKQFSQMNKIKKMALRVIAERLSEEEIGGLKELFKMIDTDNSGTITFEELKAGLKRVGSELMESEIKSLMDAADIDNSGTIDYGEFLAATLHINKMEREENLVVAFSYFDKDGSGYITIDELQQACTEFGLCDTPLDDMIKEIDLDNDGKIDFSEFTAMMKKGDGVGRSRTMRNNLNFNIAEAFGVEDTSSTAKSDDSPK C709C17DFAF74B70 cdpkb_arath Calcium-dependent protein kinase 11 At1g35670 CPK11 CDPK2 F15O4.8 Calcium-dependent protein kinase isoform CDPK2 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress METKPNPRRPSNTVLPYQTPRLRDHYLLGKKLGQGQFGTTYLCTEKSTSANYACKSIPKRKLVCREDYEDVWREIQIMHHLSEHPNVVRIKGTYEDSVFVHIVMEVCEGGELFDRIVSKGHFSEREAVKLIKTILGVVEACHSLGVMHRDLKPENFLFDSPKDDAKLKATDFGLSVFYKPGQYLYDVVGSPYYVAPEVLKKCYGPEIDVWSAGVILYILLSGVPPFWAETESGIFRQILQGKLDFKSDPWPTISEAAKDLIYKMLERSPKKRISAHEALCHPWIVDEQAAPDKPLDPAVLSRLKQFSQMNKIKKMALRVIAERLSEEEIGGLKELFKMIDTDNSGTITFEELKAGLKRVGSELMESEIKSLMDAADIDNSGTIDYGEFLAATLHMNKMEREENLVAAFSYFDKDGSGYITIDELQSACTEFGLCDTPLDDMIKEIDLDNDGKIDFSEFTAMMRKGDGVGRSRTMMKNLNFNIADAFGVDGEKSDD 2DD0ED8C234EF2F7 cpk11-dil9 2 12 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 14 unspecified role unspecified role bait bait physical association physical association The prey protein was detected using catalytically inactive, consitutively active or wild type CPK11 protein as a bait. The observation that one of the yeast two hybrid prey clones encoded the first 128 amino acids is made in section 3.5 Mapping binding sites on AtDil9 cpk11-dil9-1 5 12 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait Q39016:p.Phe306Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain F A Wed May 02 14:41:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe306ala' Tue Jun 30 15:57:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe306Ala' Q39016:p.[Asp150Ala;Phe306Ala] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain D A Fri Jul 22 11:01:26 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'phe306ala,asp150ala' Wed May 02 20:00:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp150ala,phe306ala' Tue Jun 30 19:35:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp150Ala;Phe306Ala]' Q39016:p.[Asp171Ala;Phe306Ala] mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain F A certain certain sequence position certain certain certain sequence position certain D A Wed May 02 16:30:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp171ala,phe306ala' Tue Jun 30 16:57:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp171Ala;Phe306Ala]' physical association physical association The authors state that the interacted strongly with the mutated forms of CPBK11 but weakly with the wild type CPK11 protein . Figure 1 C dil9-cpk11 6 14 unspecified role unspecified role prey prey 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 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait region 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 2 cpk11-dil9-2 10 14 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role neutral component neutral component n terminal Dsred2 protein fluorescent prot tag fluorescent protein tag fluorescent prot tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization Figure 3C The proteins colocalised to the nucleus cpk4-dil9 5 12 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait physical association physical association A consitutively active mutant form of CPK4 was used as a bait in this assay. Figure 4 cpk12-dil9 5 11 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association A consitutively active mutant form of CPK4 was used as a bait in this assay. Figure 4 The protein is named CPK9 in the UniProt database but CPK12 in TAIR database. The authors state that CPK12 interacted weakly with Dil9 since under high stringency (5mM 3-AT) the interaction did not occur cpk11-dil9-3 5 14 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey 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 5B A constitutively active mutant CPK11 protein was used as a bait in this assay cpk11-dil9-4 1 14 enzyme enzyme unspecified role unspecified role 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 12 enzyme target enzyme target substrate substrate unspecified role unspecified role region 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 region experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain thr109 phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) certain certain sequence position certain certain certain sequence position certain ser107 phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) certain certain sequence position certain certain certain sequence position certain phosphorylation phosphorylation reaction phosphorylation Figure 6B The authors state that the region used to determine phosphroylation sites was 100 to 110 amino acid residues of the Dil9 protein, whereas the UniProt sequence shows this region to be 94 to 104 amino acid residues. The authors state that one or both Thr109 and Ser107 are the preferred sites of phosphorylation Km = 25 micromolar cdpkb-di191 8 12 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait phe306ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits cdpkb-di191-1 8 12 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait asp171ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain phe306ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits cdpkb-di191-2 8 12 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait asp150ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain phe306ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits cdpkb-di191-3 4 14 unspecified role unspecified role bait bait Q39016:p.Phe306Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain F A Wed May 02 16:08:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe306ala' Tue Jun 30 19:02:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe306Ala' 12 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 physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits 1, 5 cdpkb-di191-4 4 12 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait asp171ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain phe306ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits 1 cdpkb-di191-5 4 14 unspecified role unspecified role bait bait Q39016:p.Asp150Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:11:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp150ala' Tue Jun 30 15:35:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp150Ala' Q39016:p.Phe306Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain F A Wed May 02 15:28:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe306ala' Tue Jun 30 17:28:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe306Ala' 12 unspecified role unspecified role prey prey physical association physical association We identified five clonesvcorresponding to the AtDi19 gene (Dehydration-induced 19,b At1g56280) [9]. Moreover, AtDi19 was detected in the screens performed using the three different forms of AtCPK11 baits 1 cdpkb-di191-6 3 12 unspecified role unspecified role bait bait region 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 ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Recombinant AtCPK11 interacts with AtDi19 in vitro 2 cdpkb-di191-7 9 12 unspecified role unspecified role neutral component neutral component c-terminal Dsred2 protein tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 14 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Finally, the common localization pattern of both proteinfusions (Fig. 3C) is also consistent with interactions occurring between AtCPK11 and AtDi19 in the nucleus, where both AtCPK11 and AtDi19 are present. 3 cdpk4-di191-1 4 12 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait phe305ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain physical association physical association The interaction of AtCPK4-F305A with AtDi19 is not surprising, as AtCPK11 and AtCPK4 are two of the most closely related CDPKs in Arabidopsis. 4 cpk12-di191-1 4 11 unspecified role unspecified role bait bait phe302ala experimental feature experimental feature certain certain sequence position certain certain certain sequence position certain 12 unspecified role unspecified role prey prey physical association physical association The weak interaction detected between AtCPK12-F302A and AtDi19 is consistent with the above observations as this kinase is the next most closely related member of the CDPK superfamily. 4 cdpkb-di191-10 7 14 enzyme enzyme neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q39016:p.Asp150Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain D A Wed May 02 17:47:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp150ala' Tue Jun 30 17:44:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp150Ala' ecoli Escherichia coli (strain K12) 12 enzyme target enzyme target substrate substrate neutral component neutral component region 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 ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Recombinant AtCPK11 phosphorylates AtDi19 in vitro phosphorylation of Q39083 6 Autophosphorylation and substrate phosphorylation were Ca+2-dependent Substrate phosphorylation was Ca+2-dependent cdpkb-1 7 14 putative self putative self putative self putative self region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q39016:p.Asp150Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain D A Wed May 02 14:28:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp150ala' Tue Jun 30 15:43:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp150Ala' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation true Autophosphorylation of 6xHis�AtCPK11 was more evident when it was incubated with the GST�AtDi19 fusion protein than when 6xHis�AtCPK11 was incubated alone or with GST (Fig. 6B). 6 Autophosphorylation was Ca+2-dependent and increased if Q39083 was present. autophosphorylation of Q39016 cdpkb-di191-12 7 14 enzyme enzyme neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 12 enzyme target enzyme target substrate substrate neutral component neutral component Q39083:p.Ser107Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S A Wed May 02 17:32:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser107ala' Tue Jun 30 16:41:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser107Ala' region 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 region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Q39083:p.Thr105Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain T A Wed May 02 14:28:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr105ala' Tue Jun 30 19:47:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr105Ala' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation AtDi19 bipartite NLS is phosphorylated by AtCPK11. (A) in vitro phosphorylation of the S100GTNSTLSLLR110 peptide and effect of mutation of specific amino acids on the phosphorylation of the peptide by AtCPK11. 7 phosphorylation of Q39083 peptide. Thr 105 and Ser 107 appear to be the sites of phosphorylation. Km = 25 micromolar Thr 105 and Ser 107 appear to be the sites of phosphorylation as mutations of these residues reduced phosphorylation of the peptide to only 30% and 23% relative to the wild type, respectively