scholarly journals The nitrogen phosphotransferase regulator PtsN (EIIANtr) regulates inorganic polyphosphate production in Escherichia coli

2021 ◽  
Author(s):  
Marvin Q. Bowlin ◽  
Abagail R Long ◽  
Joshua T Huffines ◽  
Michael Jeffrey Gray
Keyword(s):  
Escherichia Coli ◽  
Regulatory Networks ◽  
Stress Condition ◽  
Inorganic Polyphosphate ◽  
Promote Cell Survival ◽  
Substantial Progress ◽  
The Media ◽  
Stress Signals ◽  
Stress Sensing

Inorganic polyphosphate (polyP) is synthesized by bacteria under stressful environmental conditions and acts by a variety of mechanisms to promote cell survival. While the kinase that synthesizes polyP (PPK, enocoded by the ppk gene) is well known, little is understood about how environmental stress signals lead to activation of this enzyme. Previous work has shown that the transcriptional regulators DksA, RpoN (σ54), and RpoE (σ24) positively regulate polyP production, but not ppk transcription, in Escherichia coli. In this work, we set out to examine the role of the alternative sigma factor RpoN and nitrogen starvation stress response pathways in controlling polyP synthesis in more detail. In the course of these experiments, we identified GlnG, GlrR, PhoP, PhoQ, RapZ, and GlmS as proteins that affect polyP production, and uncovered a central role for the nitrogen phosphotransferase regulator PtsN (EIIANtr) in a polyP regulatory pathway, acting upstream of DksA, downstream of RpoN, and apparently independently of RpoE. However, none of these regulators appears to act directly on PPK, and the mechanism(s) by which they modulate polyP production remain unclear. Unexpectedly, we also found that the pathways that regulate polyP production vary depending not only on the stress condition applied, but also on the composition of the media in which the cells were grown before exposure to polyP-inducing stress. These results constitute substantial progress towards deciphering the regulatory networks driving polyP production under stress, but highlight the remarkable complexity of this regulation and its connections to a broad range of stress-sensing pathways.

scholarly journals Chaperone-Mediated Stress Sensing in Mycobacterium tuberculosis Enables Fast Activation and Sustained Response

mSystems ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Satyajit D. Rao ◽  
Pratik Datta ◽  
Maria Laura Gennaro ◽  
Oleg A. Igoshin
Keyword(s):  
Stress Response ◽  
Surface Stress ◽  
Regulatory Networks ◽  
Content Type ◽  
Chaperone Dnak ◽  
Model Predictions ◽  
Dynamical Properties ◽  
Fast Activation ◽  
Stress Sensing

ABSTRACT Dynamical properties of gene regulatory networks are tuned to ensure bacterial survival. In mycobacteria, the MprAB-σE network responds to the presence of stressors, such as surfactants that cause surface stress. Positive feedback loops in this network were previously predicted to cause hysteresis, i.e., different responses to identical stressor levels for prestressed and unstressed cells. Here, we show that hysteresis does not occur in nonpathogenic Mycobacterium smegmatis but does occur in Mycobacterium tuberculosis. However, the observed rapid temporal response in M. tuberculosis is inconsistent with the model predictions. To reconcile these observations, we implement a recently proposed mechanism for stress sensing, namely, the release of MprB from the inhibitory complex with the chaperone DnaK upon the stress exposure. Using modeling and parameter fitting, we demonstrate that this mechanism can accurately describe the experimental observations. Furthermore, we predict perturbations in DnaK expression that can strongly affect dynamical properties. Experiments with these perturbations agree with model predictions, confirming the role of DnaK in fast and sustained response. IMPORTANCE Gene regulatory networks controlling stress response in mycobacterial species have been linked to persistence switches that enable bacterial dormancy within a host. However, the mechanistic basis of switching and stress sensing is not fully understood. In this paper, combining quantitative experiments and mathematical modeling, we uncover how interactions between two master regulators of stress response—the MprAB two-component system (TCS) and the alternative sigma factor σE—shape the dynamical properties of the surface stress network. The result show hysteresis (history dependence) in the response of the pathogenic bacterium M. tuberculosis to surface stress and lack of hysteresis in nonpathogenic M. smegmatis. Furthermore, to resolve the apparent contradiction between the existence of hysteresis and fast activation of the response, we utilize a recently proposed role of chaperone DnaK in stress sensing. These result leads to a novel system-level understanding of bacterial stress response dynamics.

scholarly journals Interactions between DksA and Stress-Responsive Alternative Sigma Factors Control Inorganic Polyphosphate Accumulation in Escherichia coli

2020 ◽  
Vol 202 (14) ◽  
Author(s):  
Michael J. Gray
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Nutrient Limitation ◽  
Protein Function ◽  
Regulatory Protein ◽  
Sigma Factors ◽  
Inorganic Polyphosphate ◽  
Content Type ◽  
Alternative Sigma Factors

ABSTRACT Bacteria synthesize inorganic polyphosphate (polyP) in response to a variety of different stress conditions. polyP protects bacteria by acting as a protein-stabilizing chaperone, metal chelator, or regulator of protein function, among other mechanisms. However, little is known about how stress signals are transmitted in the cell to lead to increased polyP accumulation. Previous work in the model enterobacterium Escherichia coli has indicated that the RNA polymerase-binding regulatory protein DksA is required for polyP synthesis in response to nutrient limitation stress. In this work, I set out to characterize the role of DksA in polyP regulation in more detail. I found that overexpression of DksA increases cellular polyP content (explaining the long-mysterious phenotype of dksA overexpression rescuing growth of a dnaK mutant at high temperatures) and characterized the roles of known functional residues of DksA in this process, finding that binding to RNA polymerase is required but that none of the other functions of DksA appear to be necessary. Transcriptomics revealed genome-wide transcriptional changes upon nutrient limitation, many of which were affected by DksA, and follow-up experiments identified complex interactions between DksA and the stress-sensing alternative sigma factors FliA, RpoN, and RpoE that impact polyP production, indicating that regulation of polyP synthesis is deeply entwined in the multifactorial stress response network of E. coli. IMPORTANCE Inorganic polyphosphate (polyP) is an evolutionarily ancient, widely conserved biopolymer required for stress resistance and pathogenesis in diverse bacteria, but we do not understand how its synthesis is regulated. In this work, I gained new insights into this process by characterizing the role of the transcriptional regulator DksA in polyP regulation in Escherichia coli and identifying previously unknown links between polyP synthesis and the stress-responsive alternative sigma factors FliA, RpoN, and RpoE.

scholarly journals Complex interactions between DksA and stress-responsive alternative sigma factors control inorganic polyphosphate accumulation in Escherichia coli

2020 ◽  
Author(s):  
Michael J. Gray
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Nutrient Limitation ◽  
Protein Function ◽  
Regulatory Protein ◽  
Sigma Factors ◽  
Inorganic Polyphosphate ◽  
Complex Interactions ◽  
Alternative Sigma Factors

ABSTRACTBacteria synthesize inorganic polyphosphate (polyP) in response to a variety of different stress conditions. PolyP protects bacteria by acting as a protein-stabilizing chaperone, metal chelator, or regulator of protein function, among other mechanisms. However, little is known about how stress signals are transmitted in the cell to lead to increased polyP accumulation. Previous work in the model enterobacterium Escherichia coli has indicated that the RNA polymerase-binding regulatory protein DksA is required for polyP synthesis in response to nutrient limitation stress. In this work, I set out to characterize the role of DksA in polyP regulation in more detail. I found that overexpression of DksA increases cellular polyP content (explaining the long-mysterious phenotype of dksA overexpression rescuing growth of a dnaK mutant at high temperature) and characterized the roles of known functional residues of DksA in this process, finding that binding to RNA polymerase is required, but none of the other functions of DksA appear to be necessary. Transcriptomics revealed genome-wide transcriptional changes upon nutrient limitation, many of which were affected by DksA, and follow-up experiments identified complex interactions between DksA and the stress-sensing alternative sigma factors FliA, RpoN, and RpoE that impact polyP production, indicating that regulation of polyP synthesis is deeply entwined in the multifactorial stress response network of E. coli.IMPORTANCEInorganic polyphosphate (polyP) is an evolutionarily ancient, widely conserved biopolymer required for stress resistance and pathogenesis in diverse bacteria, but we do not understand how its synthesis is regulated. In this work, I gained new insights into this process by characterizing the role of the transcriptional regulator DksA in polyP regulation in Escherichia coli and identifying previously unknown links between polyP synthesis and the stress-responsive alternative sigma factors FliA, RpoN, and RpoE.

scholarly journals Chaperone-mediated stress-sensing in Mycobacterium tuberculosis enables fast activation and sustained response

2020 ◽  
Author(s):  
Satyajit D. Rao ◽  
Pratik Datta ◽  
Maria Laura Gennaro ◽  
Oleg A. Igoshin
Keyword(s):  
Stress Response ◽  
Surface Stress ◽  
Regulatory Networks ◽  
Chaperone Dnak ◽  
Model Predictions ◽  
Gene Regulatory ◽  
Dynamical Properties ◽  
Fast Activation ◽  
Stress Sensing

ABSTRACTDynamical properties of gene-regulatory networks are tuned to ensure bacterial survival. In mycobacteria, MprAB-σE network responds to the presence of stressors, such as surfactants causing surface stress. Positive feedback loops in this network were previously predicted to cause hysteresis, i.e. different responses to identical stressor levels for pre-stressed and unstressed cells. Here we show that hysteresis does not occur in non-pathogenic Mycobacterium smegmatis but occurs in Mycobacterium tuberculosis. However, the observed rapid temporal response in M. tuberculosis is inconsistent with the model predictions. To reconcile these observations, we implement a recently proposed mechanism for stress-sensing: the release of MprB from the inhibitory complex with chaperone DnaK upon the stress exposure. Using modeling and parameter fitting, we demonstrate that this mechanism can accurately describe the experimental observations. Furthermore, we predict perturbations in DnaK expression that can strongly affect dynamical properties. Experiments with these perturbations agree with model predictions, confirming the role of DnaK in fast and sustained response.IMPORTANCEGene-regulatory networks controlling stress response in mycobacterial species have been linked to persistence switches enabling the bacterial dormancy within a host. However, the mechanistic basis of switching and stress sensing is not fully understood. In this paper, combining quantitative experiments and mathematical modeling, we uncover how interactions between two master regulators of stress response -- MprAB two-component system and alternative sigma factor σE – shape the dynamical properties of surface-stress network. The result show hysteresis (history dependence) in the response of pathogenic bacteria M. tuberculosis to surface stress and lack of the hysteresis in non-pathogenic M. smegmatis. Furthermore, to resolve the apparent contradiction between the existence of hysteresis and fast activation of the response, we utilize a recently proposed role of chaperone DnaK in stress-sensing. The results leads to a novel system-level understanding of bacterial stress-response dynamics.

MEDIA SOCIALIZATION OF PERSONALITY IN THE CONTEXT OF INMUTATION IN THE MASS MEDIA

2021 ◽  
pp. 104-109
Author(s):  
Chernysh O.O.
Keyword(s):  
Mass Media ◽  
Negative Impact ◽  
Effective Interaction ◽  
Individual Characteristics ◽  
Media Culture ◽  
Active Growth ◽  
Media Space ◽  
The Media ◽  
The Individual

The urgency of the researched problem is connected with the growing role of mass media in modern conditions leads to change of values and transformation of identity of the person. The active growth of the role of the media, their influence on the formation and development of personality leads to the concept of “media socialization” and immutation in the media. The aim of the study is to outline the possibilities of the process of media socialization in the context of immutation in the media. The methods of our research are: analysis of pedagogical, psychological, literature, synthesis, comparison, generalization. The article analyzes the views of domestic and foreign scientists on the problem of immutation in the media and the transformation of the information space. In the context of the mass nature of the immutation of society, the concept of “media socialization” becomes relevant, which is the basis for reducing the negative impact of the media on the individual.The author identifies the lack of a thorough study of the concept of “media socialization” in modern scientific thought. Thus, media socialization is associated with the transformation of traditional means of socialization, and is to assimilate and reproduce the social experience of mankind with the help of new media.The article analyzes the essence of the concepts “media space”, “mass media” and “immutation”. The influence of mass media on the formation and development of the modern personality is described in detail.The study concluded that it is necessary to form a media culture of the individual, to establish safe and effective interaction of young people with the modern media system, the formation of media awareness, media literacy and media competence in accordance with age and individual characteristics for successful media socialization. The role of state bodies in solving the problem of media socialization of the individual was also determined. It is determined that the process of formation of media culture in youth should take place at the level of traditional institutions of socialization of the individual.The author sees the prospect of further research in a detailed analysis and study of the potential of educational institutions as an institution and a means of counteracting the mass nature of the immutation of society.Key words: immutation, media socialization, mass media, media space, information.

Cartesian Fingerprints in Beckett's Imagination Dead Imagine

2012 ◽  
Vol 21 (2) ◽  
pp. 223-243
Author(s):  
Irit Degani-Raz
Keyword(s):  
The Self ◽  
Levels Of Abstraction ◽  
The Media ◽  
The Mind ◽  
Limiting Case

The idea that Beckett investigates in his works the limits of the media he uses has been widely discussed. In this article I examine the fiction Imagination Dead Imagine as a limiting case in Beckett's exploration of limits at large and the limits of the media he uses in particular. Imagination Dead Imagine is shown to be the self-reflexive act of an artist who imaginatively explores the limits of that ultimate medium – the artist's imagination itself. My central aim is to show that various types of structural homologies (at several levels of abstraction) can be discerned between this poetic exploration of the limits of imagination and Cartesian thought. The homologies indicated here transcend what might be termed as ‘Cartesian typical topics’ (such as the mind-body dualism, the cogito, rationalism versus empiricism, etc.). The most important homologies that are indicated here are those existing between the role of imagination in Descartes' thought - an issue that until only a few decades ago was quite neglected, even by Cartesian scholars - and Beckett's perception of imagination. I suggest the use of these homologies as a tool for tracing possible sources of inspiration for Beckett's Imagination Dead Imagine.

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