Bacterial sensor kinase TodS interacts with agonistic and antagonistic signals

被引:59
作者
Busch, Andreas
Lacal, Jesus
Martos, Ariadna
Ramos, Juan L.
Krell, Tino
机构
[1] CSIC, Estac Expt Zaidin, Dept Environm Protect, E-18008 Granada, Spain
[2] CSIC, Ctr Invest Biol, E-28040 Madrid 9, Spain
关键词
histidine kinases; isothermal titration calorimetry; Pseudomonas; two-component systems; aromatic hydrocarbons;
D O I
10.1073/pnas.0701547104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The TodS/TodT two-component system controls expression of the toluene dioxygenase (TOD) pathway for the metabolism of toluene in Pseudomonas putida DOT-T1E. TodS is a sensor kinase that ultimately controls tod gene expression through its cognate response regulator, TodT. We used isothermal titration calorimetry to study the binding of different compounds to TodS and related these findings to their capacity to induce gene expression in vivo. Agonistic compounds bound to TodS and induced gene expression in vivo. Toluene was a powerful agonist, but ortho-substitutions of toluene reduced or abolished in vivo responses, although TodS recognized o-xylene with high affinity. These compounds were called antagonists. We show that agonists and antagonists compete for binding to TodS both in vitro and in vivo. The failure of antagonists to induce gene expression in vivo correlated with their inability to stimulate TodS autophosphorylation in vitro. We propose intramolecular TodS signal transmission, not molecular recognition of compounds by TodS, to be the phenomenon that determines whether a given compound will lead to activation of expression of the tod genes. Molecular modeling identified residues F46, 174, F79, and 1114 as being potentially involved in the binding of effector molecules. Alanine substitution mutants of these residues reduced affinities (2- to 345-fold) for both agonistic and antagonistic compounds. Our data indicate that determining the inhibitory activity of antagonists is a potentially fruitful alternative to design specific two-component system inhibitors for the development of new drugs to inhibit processes regulated by two-component systems.
引用
收藏
页码:13774 / 13779
页数:6
相关论文
共 39 条
[1]   Survey of the number of two-component response regulator genes in the complete and annotated genome sequences of prokaryotes [J].
Ashby, MK .
FEMS MICROBIOLOGY LETTERS, 2004, 231 (02) :277-281
[2]   Recognition of antimicrobial peptides by a bacterial sensor kinase [J].
Bader, MW ;
Sanowar, S ;
Daley, ME ;
Schneider, AR ;
Cho, US ;
Xu, WQ ;
Klevit, RE ;
Le Moual, H ;
Miller, S .
CELL, 2005, 122 (03) :461-472
[3]   Regulation of Salmonella typhimurium virulence gene expression by cationic antimicrobial peptides [J].
Bader, MW ;
Navarre, WW ;
Shiau, W ;
Nikaido, H ;
Frye, JG ;
McClelland, M ;
Fang, FC ;
Miller, SI .
MOLECULAR MICROBIOLOGY, 2003, 50 (01) :219-230
[4]  
Bates PA, 2001, PROTEINS, P39
[5]   Analysis of the π-π stacking interactions between the aminoglycoside antibiotic kinase APH(3′)-IIIa and its nucleotide ligands [J].
Boehr, DD ;
Farley, AR ;
Wright, GD ;
Cox, JR .
CHEMISTRY & BIOLOGY, 2002, 9 (11) :1209-1217
[6]   Structural basis of m7GpppG binding to the nuclear cap-binding protein complex [J].
Calero, G ;
Wilson, KF ;
Ly, T ;
Rios-Steiner, JL ;
Clardy, JC ;
Cerione, RA .
NATURE STRUCTURAL BIOLOGY, 2002, 9 (12) :912-917
[7]   Two-component signal transduction systems, environmental signals, and virulence [J].
Calva, E ;
Oropeza, R .
MICROBIAL ECOLOGY, 2006, 51 (02) :166-176
[8]   Metal bridges between the PhoQ sensor domain and the membrane regulate transmembrane signaling [J].
Cho, US ;
Bader, MW ;
Amaya, MF ;
Daley, ME ;
Klevit, RE ;
Miller, SI ;
Xu, WQ .
JOURNAL OF MOLECULAR BIOLOGY, 2006, 356 (05) :1193-1206
[9]   Identification and sequence analysis of two regulatory genes involved in anaerobic toluene metabolism by strain T1 [J].
Coschigano, PW ;
Young, LY .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1997, 63 (02) :652-660
[10]   A fully automatic evolutionary classification of protein folds: Dali Domain Dictionary version 3 [J].
Dietmann, S ;
Park, J ;
Notredame, C ;
Heger, A ;
Lappe, M ;
Holm, L .
NUCLEIC ACIDS RESEARCH, 2001, 29 (01) :55-57