Functional Characterization in Vitro of All Two-component Signal Transduction Systems from Escherichia coli Export

@article{citeulike:4860960, abstract = {Bacteria possess a signal transduction system, referred to as a two-component system, for adaptation to external stimuli. Each two-component system consists of a sensor protein-histidine kinase (HK) and a response regulator (RR), together forming a signal transduction pathway via histidyl-aspartyl phospho-relay. A total of 30 sensor HKs, including as yet uncharacterized putative HKs (BaeS, BasS, CreC, CusS, HydH, RstB, YedV, and YfhK), and a total of 34 RRs, including putative RRs (BaeR, BasR, CreB, CusR, HydG, RstA, YedW, YfhA, YgeK, and YhjB), have been suggested to exist in Escherichia coli. We have purified the carboxyl-terminal catalytic domain of 27 sensor HKs and the full-length protein of all 34 RRs to apparent homogeneity. Self-phosphorylation in vitro was detected for 25 HKs. The rate of self-phosphorylation differed among HKs, whereas the level of phosphorylation was generally co-related with the phosphorylation rate. However, the phosphorylation level was low for ArcB, HydH, NarQ, and NtrB even though the reaction rate was fast, whereas the level was high for the slow phosphorylation species BasS, CheA, and CreC. By using the phosphorylated HKs, we examined trans-phosphorylation in vitro of RRs for all possible combinations. Trans-phosphorylation of presumed cognate RRs by HKs was detected, for the first time, for eight pairs, BaeS-BaeR, BasS-BasR, CreC-CreB, CusS-CusR, HydH-HydG, RstB-RstA, YedV-YedW, and YfhK-YfhA. All trans-phosphorylation took place within less than 1/2 min, but the stability of phosphorylated RRs differed, indicating the involvement of de-phosphorylation control. In addition to the trans-phosphorylation between the cognate pairs, we detected trans-phosphorylation between about 3\% of non-cognate HK-RR pairs, raising the possibility that the cross-talk in signal transduction takes place between two-component systems. 10.1074/jbc.M410104200}, author = {Yamamoto, Kaneyoshi and Hirao, Kiyo and Oshima, Taku and Aiba, Hirofumi and Utsumi, Ryutaro and Ishihama, Akira}, citeulike-article-id = {4860960}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M410104200}, citeulike-linkout-1 = {http://www.jbc.org/cgi/content/abstract/280/2/1448}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/15522865}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=15522865}, day = {14}, doi = {10.1074/jbc.M410104200}, journal = {J. Biol. Chem.}, keywords = {bacteria, crosstalk, e\_coli, phobr, phopq, two-component\_system}, month = {January}, number = {2}, pages = {1448--1456}, posted-at = {2009-06-15 18:47:26}, priority = {0}, title = {Functional Characterization in Vitro of All Two-component Signal Transduction Systems from Escherichia coli}, url = {http://dx.doi.org/10.1074/jbc.M410104200}, volume = {280}, year = {2005} }

TY - JOUR ID - citeulike:4860960 L3 - citeulike-article-id:4860960 N2 - Bacteria possess a signal transduction system, referred to as a two-component system, for adaptation to external stimuli. Each two-component system consists of a sensor protein-histidine kinase (HK) and a response regulator (RR), together forming a signal transduction pathway via histidyl-aspartyl phospho-relay. A total of 30 sensor HKs, including as yet uncharacterized putative HKs (BaeS, BasS, CreC, CusS, HydH, RstB, YedV, and YfhK), and a total of 34 RRs, including putative RRs (BaeR, BasR, CreB, CusR, HydG, RstA, YedW, YfhA, YgeK, and YhjB), have been suggested to exist in Escherichia coli. We have purified the carboxyl-terminal catalytic domain of 27 sensor HKs and the full-length protein of all 34 RRs to apparent homogeneity. Self-phosphorylation in vitro was detected for 25 HKs. The rate of self-phosphorylation differed among HKs, whereas the level of phosphorylation was generally co-related with the phosphorylation rate. However, the phosphorylation level was low for ArcB, HydH, NarQ, and NtrB even though the reaction rate was fast, whereas the level was high for the slow phosphorylation species BasS, CheA, and CreC. By using the phosphorylated HKs, we examined trans-phosphorylation in vitro of RRs for all possible combinations. Trans-phosphorylation of presumed cognate RRs by HKs was detected, for the first time, for eight pairs, BaeS-BaeR, BasS-BasR, CreC-CreB, CusS-CusR, HydH-HydG, RstB-RstA, YedV-YedW, and YfhK-YfhA. All trans-phosphorylation took place within less than 1/2 min, but the stability of phosphorylated RRs differed, indicating the involvement of de-phosphorylation control. In addition to the trans-phosphorylation between the cognate pairs, we detected trans-phosphorylation between about 3% of non-cognate HK-RR pairs, raising the possibility that the cross-talk in signal transduction takes place between two-component systems. 10.1074/jbc.M410104200 IS - 2 JF - J. Biol. Chem. EP - 1456 TI - Functional Characterization in Vitro of All Two-component Signal Transduction Systems from Escherichia coli VL - 280 SP - 1448 KW - bacteria KW - crosstalk KW - e_coli KW - phobr KW - phopq KW - two-component_system AU - Yamamoto, Kaneyoshi AU - Hirao, Kiyo AU - Oshima, Taku AU - Aiba, Hirofumi AU - Utsumi, Ryutaro AU - Ishihama, Akira PY - 2005/01/14/ UR - http://dx.doi.org/10.1074/jbc.M410104200 ER -