scholarly journals Crystallization and preliminary X-ray analysis of the receiver domain of a putative response regulator, BPSL0128, fromBurkholderia pseudomallei

2012 ◽  
Vol 68 (8) ◽  
pp. 917-922
Author(s):  
Abd Ghani Abd Aziz ◽  
Svetlana E. Sedelnikova ◽  
Sergey N. Ruzheinikov ◽  
Simon Thorpe ◽  
Rahmah Mohamed ◽  
...  
Keyword(s):  
Response Regulator ◽  
X Ray ◽  
Receiver Domain

Overexpression, purification, crystallization and preliminary X-ray diffraction analysis of the receiver domain of PhoB

1998 ◽  
Vol 54 (6) ◽  
pp. 1460-1463 ◽  
Author(s):  
Maria Solà ◽  
F.-Xavier Gomis-Rüth ◽  
Alicia Guasch ◽  
Luis Serrano ◽  
Miquel Coll
Keyword(s):  
Response Regulator ◽  
Diffusion Method ◽  
X Ray Diffraction ◽  
Signal Transduction System ◽  
Hanging Drop ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
E Coli ◽  
Receiver Domain ◽  
Two Component Signal Transduction

PhoB is the response regulator of the E. coli two-component signal transduction system for phosphate regulation. It is a transcription factor that activates more than 30 genes of the pho regulon. Crystals of the receiver domain of PhoB were obtained by applying the hanging-drop vapour-diffusion method. X-ray diffraction data have been collected using synchrotron radiation to 1.88 Å resolution. The crystals belong to the orthorhombic space group P212121 with unit-cell constants a = 34.11, b = 60.42, c = 119.97 Å. The Matthews parameter suggests that PhoB crystallizes with two molecules per asymmetric unit, suggesting that activating dimerization occurs in the crystal.

Structure of the Acinetobacter baumannii PmrA receiver domain and insights into clinical mutants affecting DNA-binding and promoting colistin resistance

2021 ◽  
Author(s):  
Samantha Palethorpe ◽  
Morgan E Milton ◽  
Everett C Pesci ◽  
John Cavanagh
Keyword(s):  
Dna Binding ◽  
Acinetobacter Baumannii ◽  
Structural Information ◽  
Response Regulator ◽  
Regulatory System ◽  
Nosocomial Pathogen ◽  
Colistin Resistance ◽  
X Ray ◽  
X Ray Crystallography ◽  
Receiver Domain

Abstract Acinetobacter baumannii is an insidious emerging nosocomial pathogen that has developed resistance to all available antimicrobials, including the last resort antibiotic, colistin. Colistin resistance often occurs due to mutations in the PmrAB two component regulatory system. To better understand the regulatory mechanisms contributing to colistin resistance, we have biochemically characterized the A. baumannii PmrA response regulator. Initial DNA-binding analysis shows that A. baumannii PmrA bound to the Klebsiella pneumoniae PmrA box motif. This prompted analysis of the putative A. baumannii PmrAB regulon which indicated that the A. baumannii PmrA consensus box is 5′- HTTAAD N5 HTTAAD. Additionally, we provide the first structural information for the A. baumannii PmrA N-terminal domain through X-ray crystallography, and we present a full-length model using molecular modeling. From these studies, we were able to infer the effects of two critical PmrA mutations, PmrA::I13M and PmrA::P102R, both of which confer increased colistin resistance. Based on these data, we suggest structural and dynamic reasons for how these mutations can affect PmrA function and hence encourage resistive traits. Understanding these mechanisms will aid in the development of new targeted antimicrobial therapies.

scholarly journals Crystal structure of nonphosphorylated receiver domain of the stress response regulator RcsB fromEscherichia coli

2016 ◽  
Vol 25 (12) ◽  
pp. 2216-2224 ◽  
Author(s):  
Ekaterina V. Filippova ◽  
Zdzislaw Wawrzak ◽  
Jiapeng Ruan ◽  
Sergii Pshenychnyi ◽  
Richard M. Schultz ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Stress Response ◽  
Response Regulator ◽  
Fromescherichia Coli ◽  
Receiver Domain

scholarly journals Deciphering the activation and recognition mechanisms of Staphylococcus aureus response regulator ArlR

2019 ◽  
Vol 47 (21) ◽  
pp. 11418-11429 ◽  
Author(s):  
Zhenlin Ouyang ◽  
Fang Zheng ◽  
Jared Y Chew ◽  
Yingmei Pei ◽  
Jinhong Zhou ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Dna Binding ◽  
Crystal Structures ◽  
Response Regulator ◽  
Regulatory System ◽  
Repeat Sequence ◽  
Effector Domain ◽  
Receiver Domain ◽  
Regulation Mechanisms ◽  
Significant Conformational Change

Abstract Staphylococcus aureus ArlRS is a key two-component regulatory system necessary for adhesion, biofilm formation, and virulence. The response regulator ArlR consists of a C-terminal DNA-binding effector domain and an N-terminal receiver domain that is phosphorylated by ArlS, the cognate transmembrane sensor histidine kinase. We demonstrate that the receiver domain of ArlR adopts the canonical α5β5 response regulator assembly, which dimerizes upon activation, using beryllium trifluoride as an aspartate phosphorylation mimic. Activated ArlR recognizes a 20-bp imperfect inverted repeat sequence in the ica operon, which is involved in intercellular adhesion polysaccharide production. Crystal structures of the inactive and activated forms reveal that activation induces a significant conformational change in the β4-α4 and β5-α5-connecting loops, in which the α4 and α5 helices constitute the homodimerization interface. Crystal structures of the DNA-binding ArlR effector domain indicate that it is able to dimerize via a non-canonical β1–β2 hairpin domain swapping, raising the possibility of a new mechanism for signal transduction from the receiver domain to effector domain. Taken together, the current study provides structural insights into the activation of ArlR and its recognition, adding to the diversity of response regulation mechanisms that may inspire novel antimicrobial strategies specifically targeting Staphylococcus.

scholarly journals Crystal Structure of the Response Regulator 02 Receiver Domain, the Essential YycF Two-Component System of Streptococcus pneumoniae in both Complexed and Native States

2004 ◽  
Vol 186 (9) ◽  
pp. 2872-2879 ◽  
Author(s):  
Colin J. Bent ◽  
Neil W. Isaacs ◽  
Timothy J. Mitchell ◽  
Alan Riboldi-Tunnicliffe
Keyword(s):  
Streptococcus Pneumoniae ◽  
Active Site ◽  
Response Regulator ◽  
Thermotoga Maritima ◽  
Two Component System ◽  
Environmental Signals ◽  
Receiver Domain ◽  
Two Component ◽  
Protein Sequence Homology ◽  
Two Component Signal Transduction

ABSTRACT A variety of bacterial cellular responses to environmental signals are mediated by two-component signal transduction systems comprising a membrane-associated histidine protein kinase and a cytoplasmic response regulator (RR), which interpret specific stimuli and produce a measured physiological response. In RR activation, transient phosphorylation of a highly conserved aspartic acid residue drives the conformation changes needed for full activation of the protein. Sequence homology reveals that RR02 from Streptococcus pneumoniae belongs to the OmpR subfamily of RRs. The structures of the receiver domains from four members of this family, DrrB and DrrD from Thermotoga maritima, PhoB from Escherichia coli, and PhoP from Bacillus subtilis, have been elucidated. These domains are globally very similar in that they are composed of a doubly wound α5β5; however, they differ remarkably in the fine detail of the β4-α4 and α4 regions. The structures presented here reveal a further difference of the geometry in this region. RR02 is has been shown to be the essential RR in the gram-positive bacterium S. pneumoniae R. Lange, C. Wagner, A. de Saizieu, N. Flint, J. Molnos, M. Stieger, P. Caspers, M. Kamber, W. Keck, and K. E. Amrein, Gene 237:223-234, 1999; J. P. Throup, K. K. Koretke, A. P. Bryant, K. A. Ingraham, A. F. Chalker, Y. Ge, A. Marra, N. G. Wallis, J. R. Brown, D. J. Holmes, M. Rosenberg, and M. K. Burnham, Mol. Microbiol. 35:566-576, 2000). RR02 functions as part of a phosphotransfer system that ultimately controls the levels of competence within the bacteria. Here we report the native structure of the receiver domain of RR02 from serotype 4 S. pneumoniae (as well as acetate- and phosphate-bound forms) at different pH levels. Two native structures at 2.3 Å, phased by single-wavelength anomalous diffraction (xenon SAD), and 1.85 Å and a third structure at pH 5.9 revealed the presence of a phosphate ion outside the active site. The fourth structure revealed the presence of an acetate molecule in the active site.

The Signaling Pathway in Histidine Kinase and the Response Regulator Complex Revealed by X-ray Crystallography and Solution Scattering

2006 ◽  
Vol 362 (1) ◽  
pp. 123-139 ◽  
Author(s):  
Seiji Yamada ◽  
Shuji Akiyama ◽  
Hiroshi Sugimoto ◽  
Hideyuki Kumita ◽  
Kazuki Ito ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Histidine Kinase ◽  
Response Regulator ◽  
X Ray ◽  
X Ray Crystallography

scholarly journals Structure of the Francisella response regulator QseB receiver domain, and characterization of QseB inhibition by antibiofilm 2-aminoimidazole-based compounds

2017 ◽  
Vol 106 (2) ◽  
pp. 223-235 ◽  
Author(s):  
Morgan E. Milton ◽  
C. Leigh Allen ◽  
Erik A. Feldmann ◽  
Benjamin G. Bobay ◽  
David K. Jung ◽  
...  
Keyword(s):  
Response Regulator ◽  
Receiver Domain

scholarly journals Cross Talk Inhibition Nullified by a Receiver Domain Missense Substitution

2015 ◽  
Vol 197 (20) ◽  
pp. 3294-3306 ◽  
Author(s):  
TuAnh Ngoc Huynh ◽  
Hsia-Yin Lin ◽  
Chris E. Noriega ◽  
Alice V. Lin ◽  
Valley Stewart
Keyword(s):  
Cross Talk ◽  
Response Regulator ◽  
Bacterial Species ◽  
Sensor Response ◽  
Acetate Metabolism ◽  
Receiver Domain ◽  
Two Component ◽  
High Interaction ◽  
High Level

ABSTRACTIn two-component signal transduction, a sensor protein transmitter module controls cognate receiver domain phosphorylation. Most receiver domain sequences contain a small residue (Gly or Ala) at position T + 1 just distal to the essential Thr or Ser residue that forms part of the active site. However, some members of the NarL receiver subfamily have a large hydrophobic residue at position T + 1. Our laboratory previously isolated a NarL mutant in which the T + 1 residue Val-88 was replaced with an orthodox small Ala. This NarL V88A mutant confers a striking phenotype in which high-level target operon expression is both signal (nitrate) and sensor (NarX and NarQ) independent. This suggests that the NarL V88A protein is phosphorylated by cross talk from noncognate sources. Although cross talk was enhanced inackAnull strains that accumulate acetyl phosphate, it persisted inpta ackAdouble null strains that cannot synthesize this compound and was observed also innarL+strains. This indicates that acetate metabolism has complex roles in mediating NarL cross talk. Contrariwise, cross talk was sharply diminished in anarcB barAdouble null strain, suggesting that the encoded sensors contribute substantially to NarL V88A cross talk. Separately, the V88A substitution altered thein vitrorates of NarL autodephosphorylation and transmitter-stimulated dephosphorylation and decreased affinity for the cognate sensor, NarX. Together, these experiments show that the residue at position T + 1 can strongly influence two distinct aspects of receiver domain function, the autodephosphorylation rate and cross talk inhibition.IMPORTANCEMany bacterial species contain a dozen or more discrete sensor-response regulator two-component systems that convert a specific input into a distinct output pattern. Cross talk, the unwanted transfer of signals between circuits, occurs when a response regulator is phosphorylated inappropriately from a noncognate source. Cross talk is inhibited in part by the high interaction specificity between cognate sensor-response regulator pairs. This study shows that a relatively subtle missense change from Val to Ala nullifies cross talk inhibition, enabling at least two noncognate sensors to enforce an inappropriate output independently of the relevant input.

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