scholarly journals Networked Chemoreceptors Benefit Bacterial Chemotaxis Performance

mBio ◽  
2016 ◽  
Vol 7 (6) ◽  
Author(s):  
Vered Frank ◽  
Germán E. Piñas ◽  
Harel Cohen ◽  
John S. Parkinson ◽  
Ady Vaknin

ABSTRACTMotile bacteria use large receptor arrays to detect and follow chemical gradients in their environment. Extended receptor arrays, composed of networked signaling complexes, promote cooperative stimulus control of their associated signaling kinases. Here, we used structural lesions at the communication interface between core complexes to create anEscherichia colistrain with functional but dispersed signaling complexes. This strain allowed us to directly study how networking of signaling complexes affects chemotactic signaling and gradient-tracking performance. We demonstrate that networking of receptor complexes provides bacterial cells with about 10-fold-heightened detection sensitivity to attractants while maintaining a wide dynamic range over which receptor adaptational modifications can tune response sensitivity. These advantages proved especially critical for chemotaxis toward an attractant source under conditions in which bacteria are unable to alter the attractant gradient.IMPORTANCEChemoreceptor arrays are found in many motile bacteria. However, although our understanding of bacterial chemotaxis is quite detailed, the signaling and behavioral advantages of networked receptor arrays had not been directly studied in cells. We have recently shown that lesions in a key interface of theE. colireceptor array diminish physical connections and functional coupling between core signaling complexes while maintaining their basic signaling capacity. In this study, we exploited an interface 2 mutant to show, for the first time, that coupling between core complexes substantially enhances stimulus detection and chemotaxis performance.

2013 ◽  
Vol 76 (4) ◽  
pp. 668-673 ◽  
Author(s):  
CHRIS TIMMONS ◽  
SHEFALI DOBHAL ◽  
JACQUELINE FLETCHER ◽  
LI MARIA MA

Foodborne illnesses caused by Salmonella enterica and Escherichia coli O157:H7 are worldwide health concerns. Rapid, sensitive, and robust detection of these pathogens in foods and in clinical and environmental samples is essential for routine food quality testing, effective surveillance, and outbreak investigations. The aim of this study was to evaluate the effect on PCR sensitivity of adding a short, AT-rich overhanging nucleotide sequence (flap) to the 5′ end of PCR primers specific for the detection of Salmonella and E. coli O157:H7. Primers targeting the invA gene of Salmonella and the rfbE gene of E. coli O157:H7 were synthesized with or without a 12-bp, AT-rich 5′ flap (5′-AATAAATCATAA-3′). Singleplex PCR, multiplex PCR, and real-time PCR sensitivity assays were conducted using purified bacterial genomic DNA and crude cell lysates of bacterial cells. The effect of background flora on detection was evaluated by spiking tomato and jalapeno pepper surface washes with E. coli O157:H7 and Salmonella Saintpaul. When targeting individual pathogens, end-point PCR assays using flap-amended primers were more efficient than nonamended primers, with 20.4 and 23.5% increases in amplicon yield for Salmonella and E. coli O157:H7, respectively. In multiplex PCR assays, a 10- to 100-fold increase in detection sensitivity was observed when the primer flap sequence was incorporated. This improvement in both singleplex and multiplex PCR efficiency and sensitivity can lead to improved Salmonella and E. coli O157:H7 detection.


2017 ◽  
Vol 114 (36) ◽  
pp. E7583-E7591 ◽  
Author(s):  
Run-Zhi Lai ◽  
Xue-Sheng Han ◽  
Frederick W. Dahlquist ◽  
John S. Parkinson

A sensory adaptation system that tunes chemoreceptor sensitivity enables motileEscherichia colicells to track chemical gradients with high sensitivity over a wide dynamic range. Sensory adaptation involves feedback control of covalent receptor modifications by two enzymes: CheR, a methyltransferase, and CheB, a methylesterase. This study describes a CheR function that opposes the signaling consequences of its catalytic activity. In the presence of CheR, a variety of mutant serine chemoreceptors displayed up to 40-fold enhanced detection sensitivity to chemoeffector stimuli. This response enhancement effect did not require the known catalytic activity of CheR, but did involve a binding interaction between CheR and receptor molecules. Response enhancement was maximal at low CheR:receptor stoichiometry and quantitative analyses argued against a reversible binding interaction that simply shifts the ON–OFF equilibrium of receptor signaling complexes. Rather, a short-lived CheR binding interaction appears to promote a long-lasting change in receptor molecules, either a covalent modification or conformation that enhances their response to attractant ligands.


Author(s):  
Sabrina Rashid ◽  
Zhicheng Long ◽  
Shashank Singh ◽  
Maryam Kohram ◽  
Harsh Vashistha ◽  
...  

The mechanisms of bacterial chemotaxis have been extensively studied for several decades, but how the physical environment influences the collective migration of bacterial cells remains less understood. Previous models of bacterial chemotaxis have suggested that the movement of migrating bacteria across obstacle-laden terrains may be slower compared with terrains without them. Here, we show experimentally that the size or density of evenly spaced obstacles do not alter the average exit rate ofEscherichia colicells from microchambers in response to external attractants, a function that is dependent on intact cell–cell communication. We also show, both by analyzing a revised theoretical model and by experimentally following single cells, that the reduced exit time in the presence of obstacles is a consequence of reduced tumbling frequency that is adjusted by theE. colicells in response to the topology of their environment. These findings imply operational short-term memory of bacteria while moving through complex environments in response to chemotactic stimuli and motivate improved algorithms for self-autonomous robotic swarms.


2012 ◽  
Vol 12 (02) ◽  
pp. 1240003 ◽  
Author(s):  
MOHAMED AL-FANDI ◽  
MOHAMMAD A. JARADAT ◽  
MOHAMMAD AL-ROUSAN ◽  
LINA AL-EBBINI ◽  
SAIED JARADAT

In this paper, we experimentally investigated the navigation system of the nonpathogenic strain of E. coli (AW405), and we developed a simulator for the locomotion performance of these swimming nanorobots. The swimming behavior of these robotic cells is sensitive to the chemical gradients in their medium. Tissue and disease cells might produce chemical signals in their surroundings. These chemicals have the potential to affect the locomotion behavior of the bacterial cells. Therefore, bacterial cells can be considered as self-navigator nanorobots that are able to discriminate between disease cells such as cancer. We exploit Bayesian decision theory as a framework in predicting the locomotion behavior of the E. coli robotic cells. Obvious agreement has been achieved between the experimental performance of our moving robotic cells and its corresponding simulation. Our current experimental and theoretical work is considered as a platform to this novel idea of early detection of problematic diseases.


2009 ◽  
Vol 390 (11) ◽  
Author(s):  
Nikita Vladimirov ◽  
Victor Sourjik

AbstractBacterial chemotaxis represents one of the simplest and best studied examples of unicellular behavior. Chemotaxis allows swimming bacterial cells to follow chemical gradients in the environment by performing temporal comparisons of ligand concentrations. The process of chemotaxis in the model bacteriumEscherichia colihas been studied in great molecular detail over the past 40 years, using a large range of experimental tools to investigate physiology, genetics and biochemistry of the system. The abundance of quantitative experimental data enabled detailed computational modeling of the pathway and theoretical analyses of such properties as robustness and signal amplification. Because of the temporal mode of gradient sensing in bacterial chemotaxis, molecular memory is an essential component of the chemotaxis pathway. Recent studies suggest that the memory time scale has been evolutionary optimized to perform optimal comparisons of stimuli while swimming in the gradient. Moreover, noise in the adaptation system, which results from variations of the adaptation rate both over time and among cells, might be beneficial for the overall chemotactic performance of the population.


2019 ◽  
Vol 35 (6) ◽  
pp. 67-72 ◽  
Author(s):  
I.V. Manukhov ◽  
L.S. Yaguzhinsky ◽  
M.V. Bermeshev ◽  
M.A. Zisman ◽  
V.G. Pevgov ◽  
...  

Toxic effect of 2-ethylnorbornane (2-ethyl(bicyclo[2.2.1]heptane) (EBH)) on bacteria has been studied using the E. coli pRecA-lux and E. coli pKatG- lux cells as lux-biosensors. It was shown that the addition of EBH to the incubation medium leads to death and growth retardation, high level oxidative stress and DNA damage in E. coli cells. It is assumed that the oxidation of EBH with atmospheric oxygen causes the formation of reactive oxygen species in the medium, which makes a major contribution to the toxicity of this substance. biosensor, luciferase, bioluminescence, inducible promoter, PrecA, PkatG The authors are grateful to Stanislav Filippovich Chalkin for the development of interdisciplinary ties in the scientific community. The work was financially supported by the Ministry of Higher Education and Science of Russia (Project Unique Identifier RFMEFI60417X0181, Agreement No. 14.604.21.0181 of 26.09.2017).


Author(s):  
Cheng Liu ◽  
Shuiqin Fang ◽  
Yachen Tian ◽  
Youxue Wu ◽  
Meijiao Wu ◽  
...  

Escherichia coli O157:H7 ( E. coli O157:H7) is a dangerous foodborne pathogen, mainly found in beef, milk, fruits, and their products, causing harm to human health or even death. Therefore, the detection of E. coli O157:H7 in food is particularly important. In this paper, we report a lateral flow immunoassay strip (LFIS) based on aggregation-induced emission (AIE) material labeling antigen as a fluorescent probe for the rapid detection of E. coli O157:H7. The detection sensitivity of the strip is 105 CFU/mL, which is 10 times higher than that of the colloidal gold test strip. This method has good specificity and stability and can be used to detect about 250 CFU of E. coli O157:H7 successfully in 25 g or 25 mL of beef, jelly, and milk. AIE-LFIS might be valuable in monitoring food pathogens for rapid detection.


2020 ◽  
Vol 3 (1) ◽  
Author(s):  
C. Keith Cassidy ◽  
Benjamin A. Himes ◽  
Dapeng Sun ◽  
Jun Ma ◽  
Gongpu Zhao ◽  
...  

AbstractTo enable the processing of chemical gradients, chemotactic bacteria possess large arrays of transmembrane chemoreceptors, the histidine kinase CheA, and the adaptor protein CheW, organized as coupled core-signaling units (CSU). Despite decades of study, important questions surrounding the molecular mechanisms of sensory signal transduction remain unresolved, owing especially to the lack of a high-resolution CSU structure. Here, we use cryo-electron tomography and sub-tomogram averaging to determine a structure of the Escherichia coli CSU at sub-nanometer resolution. Based on our experimental data, we use molecular simulations to construct an atomistic model of the CSU, enabling a detailed characterization of CheA conformational dynamics in its native structural context. We identify multiple, distinct conformations of the critical P4 domain as well as asymmetries in the localization of the P3 bundle, offering several novel insights into the CheA signaling mechanism.


2021 ◽  
Vol 2021 (11) ◽  
pp. pdb.prot101212 ◽  
Author(s):  
Michael R. Green ◽  
Joseph Sambrook

This protocol describes a convenient method for the preparation, use, and storage of competent Escherichia coli. The reported transformation efficiency of this method is ∼5 × 107 transformants/µg of plasmid DNA.


Microbiology ◽  
2021 ◽  
Vol 167 (3) ◽  
Author(s):  
Sathi Mallick ◽  
Shanti Kiran ◽  
Tapas Kumar Maiti ◽  
Anindya S. Ghosh

Escherichia coli low-molecular-mass (LMM) Penicillin-binding proteins (PBPs) help in hydrolysing the peptidoglycan fragments from their cell wall and recycling them back into the growing peptidoglycan matrix, in addition to their reported involvement in biofilm formation. Biofilms are external slime layers of extra-polymeric substances that sessile bacterial cells secrete to form a habitable niche for themselves. Here, we hypothesize the involvement of Escherichia coli LMM PBPs in regulating the nature of exopolysaccharides (EPS) prevailing in its extra-polymeric substances during biofilm formation. Therefore, this study includes the assessment of physiological characteristics of E. coli CS109 LMM PBP deletion mutants to address biofilm formation abilities, viability and surface adhesion. Finally, EPS from parent CS109 and its ΔPBP4 and ΔPBP5 mutants were purified and analysed for sugars present. Deletions of LMM PBP reduced biofilm formation, bacterial adhesion and their viability in biofilms. Deletions also diminished EPS production by ΔPBP4 and ΔPBP5 mutants, purification of which suggested an increased overall negative charge compared with their parent. Also, EPS analyses from both mutants revealed the appearance of an unusual sugar, xylose, that was absent in CS109. Accordingly, the reason for reduced biofilm formation in LMM PBP mutants may be speculated as the subsequent production of xylitol and a hindrance in the standard flow of the pentose phosphate pathway.


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