scholarly journals A universal stress protein in Mycobacterium smegmatis sequesters the cAMP-regulated lysine acyltransferase and is essential for biofilm formation

2019 ◽  
Vol 295 (6) ◽  
pp. 1500-1516 ◽  
Author(s):  
Sintu Samanta ◽  
Priyanka Biswas ◽  
Arka Banerjee ◽  
Avipsa Bose ◽  
Nida Siddiqui ◽  
...  
Keyword(s):  
Stress Responses ◽  
Mycobacterium Smegmatis ◽  
Biofilm Formation ◽  
Stress Proteins ◽  
Response Regulator ◽  
Stress Protein ◽  
Colony Morphology ◽  
Regulator Gene ◽  
Its Gene

Universal stress proteins (USPs) are present in many bacteria, and their expression is enhanced under various environmental stresses. We have previously identified a USP in Mycobacterium smegmatis that is a product of the msmeg_4207 gene and is a substrate for a cAMP-regulated protein lysine acyltransferase (KATms; MSMEG_5458). Here, we explored the role of this USP (USP4207) in M. smegmatis and found that its gene is present in an operon that also contains genes predicted to encode a putative tripartite tricarboxylate transporter (TTT). Transcription of the TTT-usp4207 operon was induced in the presence of citrate and tartrate, perhaps by the activity of a divergent histidine kinase-response regulator gene pair. A usp4207-deleted strain had rough colony morphology and reduced biofilm formation compared with the WT strain; however, both normal colony morphology and biofilm formation were restored in a Δusp4207Δkatms strain. We identified several proteins whose acetylation was lost in the Δkatms strain, and whose transcript levels increased in M. smegmatis biofilms along with that of USP4207, suggesting that USP4207 insulates KATms from its other substrates in the cell. We propose that USP4207 sequesters KATms from diverse substrates whose activities are down-regulated by acylation but are required for biofilm formation, thus providing a defined role for this USP in mycobacterial physiology and stress responses.

scholarly journals A Two-Component Regulator of Universal Stress Protein Expression and Adaptation to Oxygen Starvation in Mycobacterium smegmatis

2003 ◽  
Vol 185 (5) ◽  
pp. 1543-1554 ◽  
Author(s):  
Ronan O'Toole ◽  
Marjan J. Smeulders ◽  
Marian C. Blokpoel ◽  
Emily J. Kay ◽  
Kathryn Lougheed ◽  
...  
Keyword(s):  
Heat Stress ◽  
Protein Expression ◽  
Stationary Phase ◽  
Mycobacterium Smegmatis ◽  
Stress Proteins ◽  
Response Regulator ◽  
Stress Protein ◽  
Wild Type ◽  
Two Component ◽  
Oxygen Starvation

ABSTRACT We identified a response regulator in Mycobacterium smegmatis which plays an important role in adaptation to oxygen-starved stationary phase. The regulator exhibits strong sequence similarity to DevR/Rv3133c of M. tuberculosis. The structural gene is present on a multigene locus, which also encodes a sensor kinase. A devR mutant of M. smegmatis was adept at surviving growth arrest initiated by either carbon or nitrogen starvation. However, its culturability decreased several orders of magnitude below that of the wild type under oxygen-starved stationary-phase conditions. Two-dimensional gel analysis revealed that a number of oxygen starvation-inducible proteins were not expressed in the devR mutant. Three of these proteins are universal stress proteins, one of which is encoded directly upstream of devR. Another protein closely resembles a proposed nitroreductase, while a fifth protein corresponds to the α-crystallin (HspX) orthologue of M. smegmatis. None of the three universal stress proteins or nitroreductase, and a considerably lower amount of HspX was detected in carbon-starved wild-type cultures. A fusion of the hspX promoter to gfp demonstrated that DevR directs gene expression when M. smegmatis enters stationary phase brought about, in particular, by oxygen starvation. To our knowledge, this is the first time a role for a two-component response regulator in the control of universal stress protein expression has been shown. Notably, the devR mutant was 104-fold more sensitive than wild type to heat stress. We conclude that DevR is a stationary-phase regulator required for adaptation to oxygen starvation and resistance to heat stress in M. smegmatis.

scholarly journals Deletion of the rpoZ gene, encoding the ω subunit of RNA polymerase, results in pleiotropic surface-related phenotypes in Mycobacterium smegmatis

Microbiology ◽  
2006 ◽  
Vol 152 (6) ◽  
pp. 1741-1750 ◽  
Author(s):  
Renjith Mathew ◽  
Raju Mukherjee ◽  
Radhakrishnan Balachandar ◽  
Dipankar Chatterji
Keyword(s):  
Rna Polymerase ◽  
Mycobacterium Smegmatis ◽  
Biofilm Formation ◽  
Physiological Role ◽  
Wild Type ◽  
Gene Encoding ◽  
Biofilm Growth ◽  
Mutant Bacterium ◽  
Sliding Motility

The ω subunit, the smallest subunit of bacterial RNA polymerase, is known to be involved in maintaining the conformation of the β′ subunit and aiding its recruitment to the rest of the core enzyme assembly in Escherichia coli. It has recently been shown in Mycobacterium smegmatis, by creating a deletion mutation of the rpoZ gene encoding ω, that the physiological role of the ω subunit also includes providing physical protection to β′. Interestingly, the mutant had altered colony morphology. This paper demonstrates that the mutant mycobacterium has pleiotropic phenotypes including reduced sliding motility and defective biofilm formation. Analysis of the spatial arrangement of biofilms by electron microscopy suggests that the altered phenotype of the mutant arises from a deficiency in generation of extracellular matrix. Complementation of the mutant strain with a copy of the wild-type rpoZ gene integrated in the bacterial chromosome restored both sliding motility and biofilm formation to the wild-type state, unequivocally proving the role of ω in the characteristics observed for the mutant bacterium. Analysis of the cell wall composition demonstrated that the mutant bacterium had an identical glycopeptidolipid profile to the wild-type, but failed to synthesize the short-chain mycolic acids characteristic of biofilm growth in M. smegmatis.

scholarly journals Role of a GntR-Family Response Regulator LbrA in Listeria monocytogenes Biofilm Formation

PLoS ONE ◽  
2013 ◽  
Vol 8 (7) ◽  
pp. e70448 ◽  
Author(s):  
Andrew Wassinger ◽  
Lu Zhang ◽  
Erin Tracy ◽  
Robert S. Munson ◽  
Sophia Kathariou ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Response Regulator ◽  
Gntr Family

scholarly journals Citrus limonoids interfere with Vibrio harveyi cell–cell signalling and biofilm formation by modulating the response regulator LuxO

Microbiology ◽  
2011 ◽  
Vol 157 (1) ◽  
pp. 99-110 ◽  
Author(s):  
Amit Vikram ◽  
Palmy R. Jesudhasan ◽  
G. K. Jayaprakasha ◽  
Suresh D. Pillai ◽  
Bhimanagouda S. Patil
Keyword(s):  
Biofilm Formation ◽  
Furan Ring ◽  
Insect Pests ◽  
Vibrio Harveyi ◽  
Response Regulator ◽  
Cell Signalling ◽  
Plant Defence ◽  
Significant Inhibition ◽  
Regulator Gene ◽  
Cell Cell

Citrus limonoids are unique secondary metabolites, characterized by a triterpenoid skeleton with a furan ring. Studies have demonstrated beneficial health properties of limonoids. In addition, certain citrus limonoids play a role in plant defence against insect pests. In the present study, five limonoids were purified from sour orange and evaluated for their ability to inhibit cell–cell signalling. The purified limonoids were tested for their ability to interfere with cell–cell signalling and biofilm formation in Vibrio harveyi. Isolimonic acid, deacetylnomilinic acid glucoside and ichangin demonstrated significant inhibition of autoinducer-mediated cell–cell signalling and biofilm formation. Furthermore, isolimonic acid and ichangin treatment resulted in induced expression of the response regulator gene luxO. In addition, luxR promoter activity was not affected by isolimonic acid or ichangin. Therefore, the ability of isolimonic acid and ichangin to interfere with cell–cell signalling and biofilm formation seems to stem from the modulation of luxO expression. The results suggest that isolimonic acid and ichangin are potent modulators of bacterial cell–cell signalling.

scholarly journals Universal stress protein Rv2624c alters abundance of arginine and enhances intracellular survival by ATP binding in mycobacteria

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Qiong Jia ◽  
Xinling Hu ◽  
Dawei Shi ◽  
Yan Zhang ◽  
Meihao Sun ◽  
...  
Keyword(s):  
Mycobacterium Smegmatis ◽  
Metabolic Pathways ◽  
Biochemical Analysis ◽  
Stress Proteins ◽  
Stress Protein ◽  
Human Monocyte ◽  
Dependent Manner ◽  
Proline Metabolism ◽  
Universal Stress Protein ◽  
Induced Proteins

Abstract The universal stress protein family is a family of stress-induced proteins. Universal stress proteins affect latency and antibiotic resistance in mycobacteria. Here, we showed that Mycobacterium smegmatis overexpressing M. tuberculosis universal stress protein Rv2624c exhibits increased survival in human monocyte THP-1 cells. Transcriptome analysis suggested that Rv2624c affects histidine metabolism, and arginine and proline metabolism. LC-MS/MS analysis showed that Rv2624c affects the abundance of arginine, a modulator of both mycobacteria and infected THP-1 cells. Biochemical analysis showed that Rv2624c is a nucleotide-binding universal stress protein, and an Rv2624c mutant incapable of binding ATP abrogated the growth advantage in THP-1 cells. Rv2624c may therefore modulate metabolic pathways in an ATP-dependent manner, changing the abundance of arginine and thus increasing survival in THP-1 cells.

Retinoic acid stimulates the synthesis of a novel heat shock protein in the regenerating forelimb of the newt

1993 ◽  
Vol 71 (1-2) ◽  
pp. 43-50 ◽  
Author(s):  
Robert L. Carlone ◽  
Robert P. Boulianne ◽  
K. Marion Vijh ◽  
Heather Karn ◽  
Gordon A. D. Fraser
Keyword(s):  
Retinoic Acid ◽  
Heat Shock ◽  
Stress Proteins ◽  
Limb Regeneration ◽  
Stress Protein ◽  
Cross Reactivity ◽  
Hsp 70 ◽  
Peptide Map ◽  
Shock Proteins

Morphogenetic effects of retinoic acid (RA) on the urodele amphibian limb regenerate pattern have been well documented, but little is known regarding the mechanism of this action of RA at the molecular level. Since exogenous RA, at concentrations sufficient to cause proximalization, represents a significant stress to newts and has been shown previously to elicit increased synthesis of heat shock proteins (HSPs) in mouse embryo limb buds, we investigated the effects of this putative morphogen on the synthesis of members of the 70-kilodalton (70-kDa) stress protein family in amputated forelimbs of the newt Notophthalmus viridescens. Injection (i.p.) of RA in dimethyl sulfoxide (DMSO), at a dose sufficient to cause significant proximal–distal reduplication of the pattern in 50% of animals treated, resulted in increased synthesis and accumulation of a 73-kDa protein with a pi of approximately 6.75. The synthesis of this same protein is increased in limb tissues as a result of a brief 35 °C heat shock. This protein is electrophoretically distinct from the newt HSP 70 family members, displays a different partial peptide map, and shows no immunological cross-reactivity with an anti-human HSP 70 monoclonal antibody. It may be a member of a separate family of 70- to 73-kDa HSPs. Interestingly, the synthesis of this protein is increased and it is more abundant in control, proximal moderate-early bud stage regenerates at 6 days after i.p. injection of DMSO than in similarly treated distal regenerates. This protein is, in addition, increased in distal regenerates to proximal levels by a prior injection of RA. The significance of these findings with regard to the possible role of stress proteins in the morphogenetic processes underlying limb regeneration is discussed.Key words: heat shock, limb regeneration, retinoic acid, pattern formation, newt.

scholarly journals Biofilm Growth By Listeria Monocytogenes On Stainless Steel and Expression of Biofilm-Related Genes Under Stressing Conditions

2021 ◽  
Author(s):  
Danilo Augusto Lopes da Silva ◽  
Rafaela de Melo Tavares ◽  
Anderson Carlos Camargo ◽  
Ricardo Seiti Yamatogi ◽  
Elaine Cristina Pereira De Martinis ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Stress Responses ◽  
Biofilm Formation ◽  
Gene Networks ◽  
Stop Codon ◽  
Regulatory Gene ◽  
Premature Stop Codon ◽  
Biofilm Growth ◽  
Adverse Conditions

Abstract This research was carried out to assess the ability of L. monocytogenes for adhesion and growth in biofilm on stainless steel coupons under different stressing conditions (NaCl, curing salts and quaternary ammonium compounds - QAC), besides determining the expression of different genes involved in biofilm formation and stress response. Results from crystal violet assay revealed that one isolate carrying a premature stop codon (PMSC) in agrC gene formed high-density biofilms in the presence of QAC or cure salts (7.5% and 10%). Reverse Transcriptase-qPCR results revealed that isolates of L. monocytogenes lineages I and II presented differences in transcriptional profile of genes related to biofilm formation and adaptation to environmental conditions. In conclusion, our results demonstrated how L. monocytogenes can survive, multiply and form biofilm under adverse conditions related to food processing environments. Differences in transcriptional expression were observed, highlighting the role of regulatory gene networks for particular serotypes under different stress responses.

scholarly journals Cellular L-arginine pools modulate c-di-GMP turnover and biofilm formation in Pseudomonas putida

2020 ◽  
Author(s):  
Laura Barrientos-Moreno ◽  
María Antonia Molina-Henares ◽  
María Isabel Ramos-González ◽  
Manuel Espinosa-Urgel
Keyword(s):  
Pseudomonas Putida ◽  
Biofilm Formation ◽  
Response Regulator ◽  
Second Messenger ◽  
Colony Morphology ◽  
Diguanylate Cyclase ◽  
Arginine Biosynthesis ◽  
Amino Acid Transport System ◽  
Metabolic Signal ◽  
Biofilm Development

<p>The intracellular second messenger cyclic diguanylate (c-di-GMP) is broadly conserved in bacteria, where it influences processes such as virulence, stress resistance and biofilm development. In the plant-beneficial bacterium <em>Pseudomonas putida</em> KT2440, the response regulator with diguanylate cyclase activity CfcR is the main contributor to c-di-GMP levels in the stationary phase of growth. When overexpressed, CfcR increases c-di-GMP levels and gives rise to a pleiotropic phenotype that includes enhanced biofilm formation and crinkly colony morphology. Our group has previously reported that insertion mutants in <em>argG</em> and <em>argH</em>, the genes that encode the last two enzymes in the arginine biosynthesis pathway, do not display the crinkly colony morphology phenotype and show decreased c-di-GMP levels even in the presence of <em>cfcR</em> in multicopy (Ramos-González, M.I. <em>et al.</em> 2016. Front. Microbiol. 7, 1093). Here we present results indicating that L-arginine acts both as an environmental and as a metabolic signal that influences the lifestyles of <em>P. putida</em> through the modulation of c-di-GMP levels and changes in the expression of structural elements of biofilms. Exogenous L-arginine partially restores c-di-GMP levels in arginine biosynthesis mutants, a response that is transduced through CfcR and possibly (an)other diguanylate cyclase(s). At least three periplasmic binding proteins, each forming part of an amino acid transport system, contribute in different ways to the response to external L-arginine. We propose that the turnover of the second messenger c-di-GMP is modulated by the state of global arginine pools in the cell resulting both from anabolism and from uptake.</p>

scholarly journals Role of SCFAs for Fimbrillin-Dependent Biofilm Formation of Actinomyces oris

2018 ◽  
Vol 6 (4) ◽  
pp. 114 ◽  
Author(s):  
Itaru Suzuki ◽  
Takehiko Shimizu ◽  
Hidenobu Senpuku
Keyword(s):  
Stress Responses ◽  
Biofilm Formation ◽  
Short Chain Fatty Acids ◽  
Flow Cell ◽  
Cell System ◽  
Oral Bacteria ◽  
Microtiter Plates ◽  
Metabolic Products

Actinomyces oris expresses type 1 and 2 fimbriae on the cell surface. Type 2 fimbriae mediate co-aggregation and biofilm formation and are composed of the shaft fimbrillin FimA and the tip fimbrillin FimB. Short-chain fatty acids (SCFAs) are metabolic products of oral bacteria, but the effects of exogenous SCFAs on FimA-dependent biofilm formation are poorly understood. We performed two types of biofilm formation assays using A. oris MG1 or MG1.ΔfimA to observe the effects of SCFAs on FimA-dependent biofilm formation in 96-well and six-well microtiter plates and a flow cell system. SCFAs did not induce six- and 16-hour biofilm formation of A. oris MG1 and MG1.ΔfimA in saliva-coated 96-well and six-well microtiter plates in which metabolites produced during growth were not excluded. However, 6.25 mM butyric acid and 3.125 mM propionic acid induced FimA-dependent biofilm formation and cell death in a flow cell system in which metabolites produced during growth were excluded. Metabolites produced during growth may lead to disturbing effects of SCFAs on the biofilm formation. The pure effects of SCFAs on biofilm formation were induction of FimA-dependent biofilm formation, but the stress responses from dead cells may regulate its effects. Therefore, SCFA may play a key role in A. oris biofilm formation.

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