scholarly journals Regulation of Proteolysis of the Stationary-Phase Sigma Factor RpoS

1998 ◽  
Vol 180 (5) ◽  
pp. 1154-1158 ◽  
Author(s):  
Yanning Zhou ◽  
Susan Gottesman
Keyword(s):  
Gene Expression ◽  
Stationary Phase ◽  
Sigma Factor ◽  
Exponential Phase ◽  
Specific Substrate ◽  
Response Regulators ◽  
Rapid Degradation ◽  
The Family ◽  
Regulation Of Activity ◽  
Sigma Factor Rpos

ABSTRACT RpoS, the stationary-phase sigma factor of Escherichia coli, is responsible for increased transcription of an array of genes when cells enter stationary phase and under certain stress conditions. RpoS is rapidly degraded during exponential phase and much more slowly during stationary phase; the resulting changes in RpoS accumulation play an important role in providing differential expression of RpoS-dependent gene expression. It has previously been shown that rapid degradation of RpoS during exponential growth depends on RssB (also called SprE and MviA), a protein with homology to the family of response regulators, and on the ClpXP protease. We find that RssB regulation of proteolysis does not extend to another ClpXP substrate, bacteriophage lambda O protein, suggesting that RssB acts on the specific substrate RpoS rather than on the protease. In addition, the activity of RpoS is down-regulated by RssB when degradation is blocked. In cells blocked for RpoS degradation by a mutation inclpP, cells devoid of RssB show a four- to fivefold-higher activity of an RpoS-dependent reporter fusion than cells overproducing RssB. Therefore, RssB allows specific environmental regulation of RpoS accumulation and may also modulate activity. The regulation of degradation provides an irreversible switch, while the regulation of activity may provide a second, presumably reversible level of control.

scholarly journals An Autoregulatory Circuit Affecting Peptide Signaling in Bacillus subtilis

1999 ◽  
Vol 181 (17) ◽  
pp. 5193-5200 ◽  
Author(s):  
Beth A. Lazazzera ◽  
Iren G. Kurtser ◽  
Ryan S. McQuade ◽  
Alan D. Grossman
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Bacillus Subtilis ◽  
Stationary Phase ◽  
Sigma Factor ◽  
Exponential Phase ◽  
Peptide Signaling ◽  
Expression Of Genes ◽  
Low Concentrations ◽  
Competence And Sporulation Factor

ABSTRACT The competence and sporulation factor (CSF) of Bacillus subtilis is an extracellular pentapeptide produced from the product of phrC. CSF has at least three activities: (i) at low concentrations, it stimulates expression of genes activated by the transcription factor ComA; at higher concentrations, it (ii) inhibits expression of those same genes and (iii) stimulates sporulation. Because the activities of CSF are concentration dependent, we measured the amount of extracellular CSF produced by cells. We found that by mid-exponential phase, CSF accumulated to concentrations (1 to 5 nM) that stimulate ComA-dependent gene expression. Upon entry into stationary phase, CSF reached 50 to 100 nM, concentrations that stimulate sporulation and inhibit ComA-dependent gene expression. Transcription of phrC was found to be controlled by two promoters: P1, which precedes rapC, the gene upstream ofphrC; and P2, which directs transcription ofphrC only. Both RapC and CSF were found to be part of autoregulatory loops that affect transcription from P1, which we show is activated by ComA∼P. RapC negatively regulates its own expression, presumably due to its ability to inhibit accumulation of ComA∼P. CSF positively regulates its own expression, presumably due to its ability to inhibit RapC activity. Transcription from P2, which is controlled by the alternate sigma factor ςH, increased as cells entered stationary phase, contributing to the increase in extracellular CSF at this time. In addition to controlling transcription ofphrC, ςH appears to control expression of at least one other gene required for production of CSF.

scholarly journals Gene expression and protein levels of the stationary phase sigma factor, RpoS, in continuously-fed Pseudomonas aeruginosa biofilms

2001 ◽  
Vol 199 (1) ◽  
pp. 67-71 ◽  
Author(s):  
Karen D. Xu ◽  
Michael J. Franklin ◽  
Chul-Ho Park ◽  
Gordon A. McFeters ◽  
Philip S. Stewart
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Stationary Phase ◽  
Sigma Factor ◽  
Protein Levels ◽  
Sigma Factor Rpos

scholarly journals Coordinated Hibernation of Transcriptional and Translational Apparatus during Growth Transition ofEscherichia colito Stationary Phase

mSystems ◽  
2018 ◽  
Vol 3 (5) ◽  
Author(s):  
Hideji Yoshida ◽  
Tomohiro Shimada ◽  
Akira Ishihama
Keyword(s):  
Stationary Phase ◽  
Sigma Factor ◽  
Exponential Phase ◽  
Screening System ◽  
Content Type ◽  
Genome Expression ◽  
70S Ribosome ◽  
K 12 ◽  
Sigma Subunit ◽  
Translational Apparatus

ABSTRACTIn the process ofEscherichia coliK-12 growth from exponential phase to stationary, marked alteration takes place in the pattern of overall genome expression through modulation of both parts of the transcriptional and translational apparatus. In transcription, the sigma subunit with promoter recognition properties is replaced from the growth-related factor RpoD by the stationary-phase-specific factor RpoS. The unused RpoD is stored by binding with the anti-sigma factor Rsd. In translation, the functional 70S ribosome is converted to inactive 100S dimers through binding with the ribosome modulation factor (RMF). Up to the present time, the regulatory mechanisms of expression of these two critical proteins, Rsd and RMF, have remained totally unsolved. In this study, attempts were made to identify the whole set of transcription factors involved in transcription regulation of thersdandrmfgenes using the newly developed promoter-specific transcription factor (PS-TF) screening system. In the first screening, 74 candidate TFs with binding activity to both of thersdandrmfpromoters were selected from a total of 194 purified TFs. After 6 cycles of screening, we selected 5 stress response TFs, ArcA, McbR, RcdA, SdiA, and SlyA, for detailed analysisin vitroandin vivoof their regulatory roles. Results indicated that bothrsdandrmfpromoters are repressed by ArcA and activated by McbR, RcdA, SdiA, and SlyA. We propose the involvement of a number of TFs in simultaneous and coordinated regulation of the transcriptional and translational apparatus. By using genomic SELEX (gSELEX) screening, each of the five TFs was found to regulate not only thersdandrmfgenes but also a variety of genes for growth and survival.IMPORTANCEDuring the growth transition ofE. colifrom exponential phase to stationary, the genome expression pattern is altered markedly. For this alteration, the transcription apparatus is altered by binding of anti-sigma factor Rsd to the RpoD sigma factor for sigma factor replacement, while the translation machinery is modulated by binding of RMF to 70S ribosome to form inactive ribosome dimer. Using the PS-TF screening system, a number of TFs were found to bind to both thersdandrmfpromoters, of which the regulatory roles of 5 representative TFs (one repressor ArcA and the four activators McbR, RcdA, SdiA, and SlyA) were analyzed in detail. The results altogether indicated the involvement of a common set of TFs, each sensing a specific environmental condition, in coordinated hibernation of the transcriptional and translational apparatus for adaptation and survival under stress conditions.

scholarly journals Unraveling the regulation of sophorolipid biosynthesis in Starmerella bombicola

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Sofie Lodens ◽  
Sophie L K W Roelants ◽  
Goedele Luyten ◽  
Robin Geys ◽  
Pieter Coussement ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stationary Phase ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Exponential Phase ◽  
Biosynthetic Gene ◽  
Reporter System ◽  
Gfp Expression ◽  
Qpcr Analysis ◽  
Starmerella Bombicola

ABSTRACT Starmerella bombicola very efficiently produces the secondary metabolites sophorolipids (SLs). Their biosynthesis is not-growth associated and highly upregulated in the stationary phase. Despite high industrial and academic interest, the underlying regulation of SL biosynthesis remains unknown. In this paper, potential regulation of SL biosynthesis through the telomere positioning effect (TPE) was investigated, as the SL gene cluster is located adjacent to a telomere. An additional copy of this gene cluster was introduced elsewhere in the genome to investigate if this results in a decoy of regulation. Indeed, for the new strain, the onset of SL production was shifted to the exponential phase. This result was confirmed by RT-qPCR analysis. The TPE effect was further investigated by developing and applying a suitable reporter system for this non-conventional yeast, enabling non-biased comparison of gene expression between the subtelomeric CYP52M1- and the URA3 locus. This was done with a constitutive endogenous promotor (pGAPD) and one of the endogenous promotors of the SL biosynthetic gene cluster (pCYP52M1). A clear positioning effect was observed for both promotors with significantly higher GFP expression levels at the URA3 locus. No clear GFP upregulation was observed in the stationary phase for any of the new strains.

scholarly journals Different Spectra of Stationary-Phase Mutations in Early-Arising versus Late-Arising Mutants of Pseudomonas putida: Involvement of the DNA Repair Enzyme MutY and the Stationary-Phase Sigma Factor RpoS

2002 ◽  
Vol 184 (24) ◽  
pp. 6957-6965 ◽  
Author(s):  
Signe Saumaa ◽  
Andres Tover ◽  
Lagle Kasak ◽  
Maia Kivisaar
Keyword(s):  
Stationary Phase ◽  
Pseudomonas Putida ◽  
Transcriptional Activation ◽  
Sigma Factor ◽  
Phenol Degradation ◽  
Test System ◽  
Growth Barriers ◽  
Mutational Processes ◽  
Sigma Factor Rpos ◽  
Selection Of

ABSTRACT Stationary-phase mutations occur in populations of stressed, nongrowing, and slowly growing cells and allow mutant bacteria to overcome growth barriers. Mutational processes in starving cells are different from those occurring in growing bacteria. Here, we present evidence that changes in mutational processes also take place during starvation of bacteria. Our test system for selection of mutants based on creation of functional promoters for the transcriptional activation of the phenol degradation genes pheBA in starving Pseudomonas putida enables us to study base substitutions (C-to-A or G-to-T transversions), deletions, and insertions. We observed changes in the spectrum of promoter-creating mutations during prolonged starvation of Pseudomonas putida on phenol minimal plates. One particular C-to-A transversion was the prevailing mutation in starving cells. However, with increasing time of starvation, the importance of this mutation decreased but the percentage of other types of mutations, such as 2- to 3-bp deletions, increased. The rate of transversions was markedly elevated in the P. putida MutY-defective strain. The occurrence of 2- to 3-bp deletions required the stationary-phase sigma factor RpoS, which indicates that some mutagenic pathway is positively controlled by RpoS in P. putida.

scholarly journals Spatial organization of different sigma factor activities and c-di-GMP signalling within the three-dimensional landscape of a bacterial biofilm

Open Biology ◽  
2018 ◽  
Vol 8 (8) ◽  
pp. 180066 ◽  
Author(s):  
Gisela Klauck ◽  
Diego O. Serra ◽  
Alexandra Possling ◽  
Regine Hengge
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Stationary Phase ◽  
Sigma Factor ◽  
Spatial Organization ◽  
Expression Patterns ◽  
Three Dimensional ◽  
Biomechanical Properties ◽  
Ribosomal Gene ◽  
Bacterial Biofilm

Bacterial biofilms are large aggregates of cells embedded in an extracellular matrix of self-produced polymers. In macrocolony biofilms of Escherichia coli , this matrix is generated in the upper biofilm layer only and shows a surprisingly complex supracellular architecture. Stratified matrix production follows the vertical nutrient gradient and requires the stationary phase σ S (RpoS) subunit of RNA polymerase and the second messenger c-di-GMP. By visualizing global gene expression patterns with a newly designed fingerprint set of Gfp reporter fusions, our study reveals the spatial order of differential sigma factor activities, stringent control of ribosomal gene expression and c-di-GMP signalling in vertically cryosectioned macrocolony biofilms. Long-range physiological stratification shows a duplication of the growth-to-stationary phase pattern that integrates nutrient and oxygen gradients. In addition, distinct short-range heterogeneity occurs within specific biofilm strata and correlates with visually different zones of the refined matrix architecture. These results introduce a new conceptual framework for the control of biofilm formation and demonstrate that the intriguing extracellular matrix architecture, which determines the emergent physiological and biomechanical properties of biofilms, results from the spatial interplay of global gene regulation and microenvironmental conditions. Overall, mature bacterial macrocolony biofilms thus resemble the highly organized tissues of multicellular organisms.

scholarly journals Characterization of a Stress-Induced Alternate Sigma Factor, RpoS, of Coxiella burnetii and Its Expression during the Development Cycle

2001 ◽  
Vol 69 (8) ◽  
pp. 4874-4883 ◽  
Author(s):  
Rekha Seshadri ◽  
James E. Samuel
Keyword(s):  
Stationary Phase ◽  
Coxiella Burnetii ◽  
Sigma Factor ◽  
Stationary Phases ◽  
Large Cell ◽  
Polyclonal Antiserum ◽  
Developmental Cycle ◽  
Growth Stages ◽  
Extracellular Environment ◽  
Sigma Factor Rpos

ABSTRACT Coxiella burnetii is an obligate intracellular bacterium that resides in an acidified phagolysosome and has a remarkable ability to persist in the extracellular environment.C. burnetii has evolved a developmental cycle that includes at least two morphologic forms, designated large cell variants (LCV) and small cell variants (SCV). Based on differential protein expression, distinct ultrastructures, and different metabolic activities, we speculated that LCV and SCV are similar to typical logarithmic- and stationary-phase growth stages. We hypothesized that the alternate sigma factor, RpoS, a global regulator of genes expressed under stationary-phase, starvation, and stress conditions in many bacteria, regulates differential expression in life cycle variants of C. burnetii. To test this hypothesis, we cloned and characterized the major sigma factor, encoded by an rpoD homologue, and the stress response sigma factor, encoded by an rpoS homologue. TherpoS gene was cloned by complementation of anEscherichia coli rpoS null mutant containing an RpoS-dependent lacZ fusion (osmY::lacZ). Expression ofC. burnetii rpoS was regulated by growth phase inE. coli (induced upon entry into stationary phase). A glutathione S-transferase–RpoS fusion protein was used to develop polyclonal antiserum against C. burnetii RpoS. Western blot analysis detected abundant RpoS in LCV but not in SCV. These results suggest that LCV and SCV are not comparable to logarithmic and stationary phases of growth and may represent a novel adaptation for survival in both the phagolysosome and the extracellular environment.

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