scholarly journals Identification of New Chromosomal Loci Involved in com Genes Expression and Natural Transformation in the Actinobacterial Model Organism Micrococcus luteus

Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1307
Author(s):  
Enzo Joaquin Torasso Kasem ◽  
Angel Angelov ◽  
Elisa Werner ◽  
Antoni Lichev ◽  
Sonja Vanderhaeghen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Natural Transformation ◽  
Micrococcus Luteus ◽  
Model Organism ◽  
Competence Development ◽  
Chemical Mutagenesis ◽  
Reporter Strain ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Genes Expression

Historically, Micrococcus luteus was one of the first organisms used to study natural transformation, one of the main routes of horizontal gene transfer among prokaryotes. However, little is known about the molecular basis of competence development in M. luteus or any other representative of the phylum of high-GC Gram-positive bacteria (Actinobacteria), while this means of genetic exchange has been studied in great detail in Gram-negative and low-GC Gram-positive bacteria (Firmicutes). In order to identify new genetic elements involved in regulation of the comEA-comEC competence operon in M. luteus, we conducted random chemical mutagenesis of a reporter strain expressing lacZ under the control of the comEA-comEC promoter, followed by the screening of dysregulated mutants. Mutants with (i) upregulated com promoter under competence-repressing conditions and (ii) mutants with a repressed com promoter under competence-inducing conditions were isolated. After genotype and phenotype screening, the genomes of several mutant strains were sequenced. A selection of putative com-influencing mutations was reinserted into the genome of the M. luteus reporter strain as markerless single-nucleotide mutations to confirm their effect on com gene expression. This strategy revealed mutations affecting com gene expression at genetic loci different from previously known genes involved in natural transformation. Several of these mutations decreased transformation frequencies by several orders of magnitude, thus indicating significant roles in competence development or DNA acquisition in M. luteus. Among the identified loci, there was a new locus containing genes with similarity to genes of the tad clusters of M. luteus and other bacteria.

scholarly journals Attenuator LRR – a regulatory tool for modulating gene expression in Gram‐positive bacteria

2021 ◽  
Author(s):  
Xia Cai ◽  
Qian Wang ◽  
Yu Fang ◽  
Die Yao ◽  
Yunda Zhan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gram Positive ◽  
Gram Positive Bacteria

scholarly journals Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions

Molecules ◽  
2019 ◽  
Vol 24 (11) ◽  
pp. 2077 ◽  
Author(s):  
Andrea Díaz-Roa ◽  
Abraham Espinoza-Culupú ◽  
Orlando Torres-García ◽  
Monamaris M. Borges ◽  
Ivan N. Avino ◽  
...  
Keyword(s):  
Micrococcus Luteus ◽  
Multidrug Resistant ◽  
Peptide Sequence ◽  
Resistant Bacteria ◽  
Gram Positive ◽  
Antimicrobial Drugs ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Conserved Domain ◽  
Health Institutions

Antibiotic resistance is at dangerous levels and increasing worldwide. The search for new antimicrobial drugs to counteract this problem is a priority for health institutions and organizations, both globally and in individual countries. Sarconesiopsis magellanica blowfly larval excretions and secretions (ES) are an important source for isolating antimicrobial peptides (AMPs). This study aims to identify and characterize a new S. magellanica AMP. RP-HPLC was used to fractionate ES, using C18 columns, and their antimicrobial activity was evaluated. The peptide sequence of the fraction collected at 43.7 min was determined by mass spectrometry (MS). Fluorescence and electronic microscopy were used to evaluate the mechanism of action. Toxicity was tested on HeLa cells and human erythrocytes; physicochemical properties were evaluated. The molecule in the ES was characterized as sarconesin II and it showed activity against Gram-negative (Escherichia coli MG1655, Pseudomonas aeruginosa ATCC 27853, P. aeruginosa PA14) and Gram-positive (Staphylococcus aureus ATCC 29213, Micrococcus luteus A270) bacteria. The lowest minimum inhibitory concentration obtained was 1.9 μM for M. luteus A270; the AMP had no toxicity in any cells tested here and its action in bacterial membrane and DNA was confirmed. Sarconesin II was documented as a conserved domain of the ATP synthase protein belonging to the Fli-1 superfamily. The data reported here indicated that peptides could be alternative therapeutic candidates for use in infections against Gram-negative and Gram-positive bacteria and eventually as a new resource of compounds for combating multidrug-resistant bacteria.

scholarly journals Hydrogen Peroxide-Mediated Killing of Caenorhabditis elegans by Streptococcus pyogenes

2002 ◽  
Vol 70 (9) ◽  
pp. 5202-5207 ◽  
Author(s):  
W. T. M. Jansen ◽  
M. Bolm ◽  
R. Balling ◽  
G. S. Chhatwal ◽  
R. Schnabel
Keyword(s):  
Hydrogen Peroxide ◽  
Caenorhabditis Elegans ◽  
Streptococcus Pyogenes ◽  
Wide Spectrum ◽  
Model Organism ◽  
Common Mechanism ◽  
Gram Positive ◽  
C Elegans ◽  
Gram Positive Bacteria ◽  
Serovar Typhimurium

ABSTRACT Caenorhabditis elegans is currently introduced as a new, facile, and cheap model organism to study the pathogenesis of gram-negative bacteria such as Pseudomonas aeruginosa and Salmonella enterica serovar Typhimurium. The mechanisms of killing involve either diffusible exotoxins or infection-like processes. Recently, it was shown that also some gram-positive bacteria kill C. elegans, although the precise mechanisms of killing remained open. We examined C. elegans as a pathogenesis model for the gram-positive bacterium Streptococcus pyogenes, a major human pathogen capable of causing a wide spectrum of diseases. We demonstrate that S. pyogenes kills C. elegans, both on solid and in liquid medium. Unlike P. aeruginosa and S. enterica serovar Typhimurium, the killing by S. pyogenes is solely mediated by hydrogen peroxide. Killing required live streptococci; the killing capacity depends on the amount of hydrogen peroxide produced, and killing can be inhibited by catalase. Major exotoxins of S. pyogenes are not involved in the killing process as confirmed by using specific toxin inhibitors and knockout mutants. Moreover, no accumulation of S. pyogenes in C. elegans is observed, which excludes the involvement of infection-like processes. Preliminary results show that S. pneumoniae can also kill C. elegans by hydrogen peroxide production. Hydrogen peroxide-mediated killing might represent a common mechanism by which gram-positive, catalase-negative pathogens kill C. elegans.

scholarly journals Resilience in the Face of Uncertainty: Sigma Factor B Fine-Tunes Gene Expression To Support Homeostasis in Gram-Positive Bacteria

2016 ◽  
Vol 82 (15) ◽  
pp. 4456-4469 ◽  
Author(s):  
Claudia Guldimann ◽  
Kathryn J. Boor ◽  
Martin Wiedmann ◽  
Veronica Guariglia-Oropeza
Keyword(s):  
Gene Expression ◽  
Sigma Factor ◽  
Environmental Changes ◽  
Regulation Of Gene Expression ◽  
Bacterial Survival ◽  
Gram Positive ◽  
Content Type ◽  
Gram Positive Bacteria ◽  
Changing Environments ◽  
The Face

ABSTRACTGram-positive bacteria are ubiquitous and diverse microorganisms that can survive and sometimes even thrive in continuously changing environments. The key to such resilience is the ability of members of a population to respond and adjust to dynamic conditions in the environment. In bacteria, such responses and adjustments are mediated, at least in part, through appropriate changes in the bacterial transcriptome in response to the conditions encountered. Resilience is important for bacterial survival in diverse, complex, and rapidly changing environments and requires coordinated networks that integrate individual, mechanistic responses to environmental cues to enable overall metabolic homeostasis. In many Gram-positive bacteria, a key transcriptional regulator of the response to changing environmental conditions is the alternative sigma factor σB. σBhas been characterized in a subset of Gram-positive bacteria, including the generaBacillus,Listeria, andStaphylococcus. Recent insight from next-generation-sequencing results indicates that σB-dependent regulation of gene expression contributes to resilience, i.e., the coordination of complex networks responsive to environmental changes. This review explores contributions of σBto resilience inBacillus,Listeria, andStaphylococcusand illustrates recently described regulatory functions of σB.

scholarly journals Small-Molecule Antibiotics Inhibiting tRNA-Regulated Gene Expression Is a Viable Strategy for Targeting Gram-Positive Bacteria

2020 ◽  
Vol 65 (1) ◽  
pp. e01247-20 ◽  
Author(s):  
Ville Y. P. Väre ◽  
Ryan F. Schneider ◽  
Haein Kim ◽  
Erica Lasek-Nesselquist ◽  
Kathleen A. McDonough ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Molecule ◽  
Bacterial Infections ◽  
Gram Positive ◽  
Content Type ◽  
T Box ◽  
Gram Positive Bacteria ◽  
Bactericidal Effects ◽  
Low Levels ◽  
Regulated Gene Expression

ABSTRACTBacterial infections and the rise of antibiotic resistance, especially multidrug resistance, have generated a clear need for discovery of novel therapeutics. We demonstrated that a small-molecule drug, PKZ18, targets the T-box mechanism and inhibits bacterial growth. The T-box is a structurally conserved riboswitch-like gene regulator in the 5′ untranslated region (UTR) of numerous essential genes of Gram-positive bacteria. T-boxes are stabilized by cognate, unacylated tRNA ligands, allowing the formation of an antiterminator hairpin in the mRNA that enables transcription of the gene. In the absence of an unacylated cognate tRNA, transcription is halted due to the formation of a thermodynamically more stable terminator hairpin. PKZ18 targets the site of the codon-anticodon interaction of the conserved stem I and reduces T-box-controlled gene expression. Here, we show that novel analogs of PKZ18 have improved MICs, bactericidal effects against methicillin-resistant Staphylococcus aureus (MRSA), and increased efficacy in nutrient-limiting conditions. The analogs have reduced cytotoxicity against eukaryotic cells compared to PKZ18. The PKZ18 analogs acted synergistically with aminoglycosides to significantly enhance the efficacy of the analogs and aminoglycosides, further increasing their therapeutic windows. RNA sequencing showed that the analog PKZ18-22 affects expression of 8 of 12 T-box controlled genes in a statistically significant manner, but not other 5′-UTR regulated genes in MRSA. Very low levels of resistance further support the existence of multiple T-box targets for PKZ18 analogs in the cell. Together, the multiple targets, low resistance, and synergy make PKZ18 analogs promising drugs for development and future clinical applications.

scholarly journals Effects of green algae and napa cabbage on life-history parameters and gut microflora of Archegozetes longisetosus (Acari: Oribatida) under laboratory conditions

2016 ◽  
Vol 53 (2) ◽  
pp. 67-78 ◽  
Author(s):  
Anna Seniczak ◽  
Anna Ligocka ◽  
Stanisław Seniczak ◽  
Zbigniew Paluszak
Keyword(s):  
Life History ◽  
Green Algae ◽  
Model Organism ◽  
Oribatid Mite ◽  
Gut Microflora ◽  
Crude Fibre ◽  
Gram Positive ◽  
Life History Parameters ◽  
Gram Positive Bacteria ◽  
Archegozetes Longisetosus

AbstractWe compared the effect of green algae (dominated byProtococcussp.) and the earlier studied napa cabbage on the life-history parameters and gut microflora of the oribatid miteArchegozetes longisetosus(a chelicerate model organism). Napa cabbage contained more crude ash, protein, and crude fibre than green algae, butA.longisetosusdeveloped better on the latter food, displaying higher fertility, lower mortality of offspring and shorter development than on napa cabbage. The gut microflora ofA.longisetosusdepended on the kind of food and developmental stage of this mite. The adults fed with napa cabbage had more abundant and more active microflora than those fed with green algae, whereas in the tritonymphs the microflora was more abundant when they were fed with green algae, and was more active in the group fed with napa cabbage. Irrespective of the treatment, the highest percentage of the isolated bacteria were gram-positive bacilli or gram-negative bacteria, but the mites feeding onProtococcussp. contained no cocci, whereas those fed with cabbage had no gram-positive bacteria.

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