scholarly journals The integrated stress response pathway activates 4E-BP to bias mRNA translation and boost antimicrobial peptide synthesis in response to bacterial infection

2017 ◽  
Author(s):  
Deepika Vasudevan ◽  
Nicholas Clark ◽  
Jessica Sam ◽  
Beatrix Ueberheide ◽  
Michael T. Marr ◽  
...  
Keyword(s):  
Bacterial Infection ◽  
Pathogenic Bacteria ◽  
Mrna Translation ◽  
Mechanism Analysis ◽  
Protein Levels ◽  
Translational Activity ◽  
Stress Response Pathway ◽  
Upstream Kinase ◽  
Transcriptional Induction

Pathogenic bacterial infection imposes considerable cellular stress on the host and often leads to attenuation of mRNA translation. In this translation-suppressive environment, it is unclear how the host synthesizes various antimicrobial peptides (AMPs) to mount innate immune response. Here, we use Drosophila as a model to demonstrate that AMP production during infection relies on a translation bias mechanism mediated by the inhibitor of cap-dependent translation 4E-BP ( Drosophila Thor), and the AMP 5′ UTRs that can undergo cap-independent translation. We found that 4E-BP is induced upon infection with the pathogenic bacteria Ecc15 by the stress-responsive transcription factor ATF4, and its upstream kinase GCN2. Moreover, loss of gcn2 , atf4 or 4e-bp compromised immunity against Ecc15 . In 4E-BP mutants, the transcriptional induction of AMPs after infection was unaffected, while the protein levels of AMPs were substantially reduced in their hemolymph. Analysis of the 5′ UTRs of AMPs using cell-based bicistronic reporters and in vitro translation analysis indicated that AMPs are translated in a cap-independent mechanism. Analysis of bicistronic reporters in the presence of 4E-BP indicate that infection enhances cap-independent translational activity associated with AMP 5′ UTRs, accounting for enhanced AMP translation during infection.

scholarly journals NSun2 promotes cell migration through methylating autotaxin mRNA

2020 ◽  
Vol 295 (52) ◽  
pp. 18134-18147
Author(s):  
Xin Xu ◽  
Yihua Zhang ◽  
Junjie Zhang ◽  
Xiaotian Zhang
Keyword(s):  
Cell Migration ◽  
Biological Effects ◽  
Mrna Translation ◽  
Glioma Cell Line ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Protein Levels ◽  
Key Factor

NSun2 is an RNA methyltransferase introducing 5-methylcytosine into tRNAs, mRNAs, and noncoding RNAs, thereby influencing the levels or function of these RNAs. Autotaxin (ATX) is a secreted glycoprotein and is recognized as a key factor in converting lysophosphatidylcholine into lysophosphatidic acid (LPA). The ATX-LPA axis exerts multiple biological effects in cell survival, migration, proliferation, and differentiation. Here, we show that NSun2 is involved in the regulation of cell migration through methylating ATX mRNA. In the human glioma cell line U87, knockdown of NSun2 decreased ATX protein levels, whereas overexpression of NSun2 elevated ATX protein levels. However, neither overexpression nor knockdown of NSun2 altered ATX mRNA levels. Further studies revealed that NSun2 methylated the 3′-UTR of ATX mRNA at cytosine 2756 in vitro and in vivo. Methylation by NSun2 enhanced ATX mRNA translation. In addition, NSun2-mediated 5-methylcytosine methylation promoted the export of ATX mRNA from nucleus to cytoplasm in an ALYREF-dependent manner. Knockdown of NSun2 suppressed the migration of U87 cells, which was rescued by the addition of LPA. In summary, we identify NSun2-mediated methylation of ATX mRNA as a novel mechanism in the regulation of ATX.

scholarly journals Safe and easy evaluation of tmRNA-SmpB-mediated trans-translation in ESKAPE pathogenic bacteria

2020 ◽  
Author(s):  
Marion Thépaut ◽  
Rodrigo Campos Da Silva ◽  
Eva Renard ◽  
Frédérique Barloy-Hubler ◽  
Eric Ennifar ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Pathogenic Bacteria ◽  
Fluorescent Protein ◽  
Translational Activity ◽  
Promising Target ◽  
A Cell ◽  
Bacterial Fitness ◽  
Green Fluorescent

AbstractBacteria cope with ribosome stalling thanks to trans-translation, a major quality control system of protein synthesis that is mediated by tmRNA, an hybrid RNA with properties of both a tRNA and an mRNA, and the small protein SmpB. Because trans-translation is absent in eukaryotes but necessary for bacterial fitness or survival, it is a promising target for the development of novel antibiotics. To facilitate screening of chemical libraries, various reliable in vitro and in vivo systems have been created for assessing trans-translational activity. However, none of these permits the safe and easy evaluation of trans-translation in pathogenic bacteria, which are obviously the ones we should be targeting. Based on green fluorescent protein (GFP) reassembly during active trans-translation, we have created a cell-free assay adapted to the rapid evaluation of trans-translation in ESKAPE bacteria, with 24 different possible combinations. It can be used for easy high-throughput screening of chemical compounds as well as for exploring the mechanism of trans-translation in these pathogens.

scholarly journals Influence of Mycobacterium bovisBacillus Calmette Guérin on In Vitro Induction of CD1 Molecules in Human Adherent Mononuclear Cells

2001 ◽  
Vol 69 (12) ◽  
pp. 7461-7470 ◽  
Author(s):  
Anna Giuliani ◽  
Salvatore P. Prete ◽  
Grazia Graziani ◽  
Angelo Aquino ◽  
Alessandra Balduzzi ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Mononuclear Cells ◽  
Gm Csf ◽  
Protein Levels ◽  
High Concentrations ◽  
Macrophage Colony ◽  
Nonpeptide Antigens ◽  
Mycobacterial Antigens ◽  
Antigen Presenting

ABSTRACT Nonpeptide antigens (including glycolipids of microbial origin) can be presented to T cells by CD1 molecules expressed on monocyte-derived dendritic cells. These HLA unrestricted responses appear to play a role in host immunity against Mycobacterium tuberculosis and other pathogenic bacteria. It is known that vaccination withMycobacterium bovis bacillus Calmette-Guérin (BCG) has limited efficacy in many clinical settings, although the reasons for its inadequacy remain unclear. Here we have investigated the influence of BCG on the induction of CD1b on human monocytes by granulocyte-macrophage colony-stimulating factor (GM-CSF), which is believed to be the principal inducer of this antigen-presenting molecule. Although BCG alone led to a slight induction of CD1b expression, this agent reduced markedly the ability of GM-CSF to induce high levels of CD1b that were typically observed in uninfected cells. Inhibition of CD1b expression in BCG-infected monocytes was apparent at both the mRNA transcript and CD1b protein levels. Down-regulation of CD1b expression by BCG was mediated, at least in part, by one or more soluble factors and could not be reversed with high concentrations of GM-CSF or a variety of other cytokines. The present results suggest that BCG could diminish the efficiency of CD1-restricted T-cell responses against nonpeptide mycobacterial antigens by reducing CD1 expression on antigen-presenting cells. These findings have potential implications for understanding the nature of the immune response elicited by BCG in humans and suggest potential strategies that could be important for the development of better vaccines for the prevention of tuberculosis.

Preliminary in Vitro Investigations on The Inhibitory Activity of The Original Dietary Supplement Oxidal® on Pathogenic Bacterial Strains

2020 ◽  
Vol 11 ◽  
pp. 37-43
Author(s):  
Prof. Teodora P. Popova ◽  
Toshka Petrova ◽  
Ignat Ignatov ◽  
Stoil Karadzhov
Keyword(s):  
Dietary Supplement ◽  
Broad Spectrum ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Clinical Effectiveness ◽  
Inhibitory Effect ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Microbial Strains

The antimicrobial action of the dietary supplement Oxidal® was tested using the classic Bauer and Kirby agar-gel diffusion method. Clinical and reference strains of Staphylococcus aureus and Escherichia coli were used in the studies. The tested dietary supplement showed a well-pronounced inhibitory effect against the microbial strains commensurable with that of the broad-spectrum chemotherapeutic agent Enrofloxacin and showed even higher activity than the broad spectrum antibiotic Thiamphenicol. The proven inhibitory effect of the tested dietary supplement against the examined pathogenic bacteria is in accordance with the established clinical effectiveness standards for antimicrobial agents.

THE ACTIVITY OF ESSENTIAL OILS OBTAINED FROM SPECIES AND INTERSPECIES HYBRIDS OF THE Mentha GENUS AGAINST SELECTED PLANT PATHOGENIC BACTERIA

2018 ◽  
Vol 17 (6) ◽  
pp. 167-174 ◽  
Author(s):  
Małgorzata Schollenberger ◽  
Tomasz M. Staniek ◽  
Elżbieta Paduch-Cichal ◽  
Beata Dasiewicz ◽  
Agnieszka Gadomska-Gajadhur ◽  
...  
Keyword(s):  
Essential Oils ◽  
Pseudomonas Syringae ◽  
Pathogenic Bacteria ◽  
Antimicrobial Effect ◽  
Mentha Piperita ◽  
Mentha Spicata ◽  
Plant Essential Oils ◽  
Plant Pathogenic Bacteria ◽  
Interspecies Hybrids

Plant essential oils of six aromatic herb species and interspecies hybrids of the family Lamiaceae – chocolate mint (Mentha piperita × ‘Chocolate’), pineapple mint (Mentha suaveolens ‘Variegata’), apple mint (Mentha × rotundifolia), spearmint (Mentha spicata), orange mint (Mentha × piperita ‘Granada’) and strawberry mint (Mentha × villosa ‘Strawberry’) – were investigated for antimicrobial effects against plant pathogenic bacteria: Agrobacterium tumefaciens, Pseudomonas syringae pv. syringae and Xanthomonas arboricola pv. corylina. The screening was carried out in vitro on agar plates filled with the target organism. All essential oils screened exhibited a higher level of antibacterial activity against A. tumefaciens and X. arboricola pv. corylina than streptomycin used as a standard in all tests. The antimicrobial effect of streptomycin and five mint oils was at the same level for P. syringae pv. syringae. There were no significant differences in the influence of the chocolate mint oil on the growth inhibition of all bacteria tested. Plant essential oils from pineapple mint, apple mint, spearmint and strawberry mint showed the weakest antimicrobial activity against P. syringae pv. syringae and the strongest towards A. tumefaciens and X. arboricola pv. corylina. The essential oils from strawberry mint, pineapple mint, spearmint and apple mint had the strongest effect on A. tumefaciens, and the lowest inhibitory activity was exhibited by the chocolate mint and orange mint essential oils. X. arboricola pv. corylina was the most sensitive to the strawberry mint, pineapple mint and spearmint oils. The chocolate mint oil showed the greatest activity against P. syringae pv. syringae.

Evaluation of Antibacterial and Antidiabetic Potential of Silver Nanoparticles using Eugenia Uniflora L. Seed Extract

2020 ◽  
Vol 13 (6) ◽  
pp. 5217-5225
Author(s):  
Guru Kumar Dugganaboyana ◽  
Chethankumar Mukunda ◽  
Suresh Darshini Inakanally
Keyword(s):  
Biomedical Applications ◽  
Pathogenic Bacteria ◽  
Seed Extract ◽  
Drug Formulation ◽  
Visible Spectroscopy ◽  
Plant Materials ◽  
X Ray ◽  
Eugenia Uniflora ◽  
Uv Visible

In recent years, green nanotechnology-based approaches using plant materials have been accepted as an environmentally friendly and cost-effective approach with various biomedical applications. In the current study, AgNPs were synthesized using the seed extract of the Eugenia uniflora L. (E.uniflora). Characterization was done using UV-Visible spectroscopy, X-ray diffraction (XRD), scanning electronic microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) analyses. The formation of AgNPs has confirmed through UV-Visible spectroscopy (at 466 nm) by the change of color owing to surface Plasmon resonance. Based on the XRD pattern, the crystalline property of AgNPs was established. The functional group existing in seed of E.uniflora extract accountable for the reduction of Ag+ ion and the stabilization of AgNPs was investigated. The morphological structures and elemental composition was determined by SEM and EDX analysis. With the growing application of AgNPs in biomedical perspectives, the biosynthesized AgNPs were evaluated for their antibacterial and along with their antidiabetic potential. The results showed that AgNPs are extremely effective with potent antidiabetic potential at a very low concentration. It also exhibited potential antibacterial activity against the three tested human pathogenic bacteria. Overall, the results highlight the effectiveness and potential applications of AgNPs in biomedical fields such as in the treatment of acute illnesses as well as in drug formulation for treating various diseases such as cancer and diabetes. It could be concluded that E. uniflora seed extract AgNPs can be used efficiently for in vitro evaluation of their antibacterial and antidiabetic effects with potent biomedical applications.

Study of Phosphocalcic Glasses from SiO2-CaO-P2O5 System with and Without Silver II. The bioactivity analysis by FTIR, SEM methods and microbiological study of silver-doped glasses

2017 ◽  
Vol 68 (6) ◽  
pp. 1188-1192
Author(s):  
Daniela Avram ◽  
Nicolae Angelescu ◽  
Dan Nicolae Ungureanu ◽  
Ionica Ionita ◽  
Iulian Bancuta ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Body Fluid ◽  
Pathogenic Bacteria ◽  
De Novo ◽  
Microbiological Examination ◽  
Doped Glasses ◽  
P2o5 System ◽  
Scanning Electron

The study in vitro of the glass powders bioactivity was performed by soaking them in simulated body fluid for 3 to 21 days at a temperature of 37�C and pH = 7.20. The synthesis de novo of hydroxyapatite, post soaking was confirmed by Fourier Transform Infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The study of the antimicrobial activity was performed by microbiological examination on two strains of pathogenic bacteria involved in postoperative nosocomial infections.

Prospective Application of Aptamer-based Assays and Therapeutics in Bloodstream Infections

2020 ◽  
Vol 20 (10) ◽  
pp. 831-840
Author(s):  
Weibin Li
Keyword(s):  
Pathogenic Bacteria ◽  
Genetic Diagnosis ◽  
Bloodstream Infections ◽  
High Specificity ◽  
Peptide Sequence ◽  
High Morbidity ◽  
Specificity And Sensitivity ◽  
Prospective Application ◽  
Small Molecule Therapeutics

Sepsis is still a severe health problem worldwide with high morbidity and mortality. Blood bacterial culture remains the gold standard for the detection of pathogenic bacteria in bloodstream infections, but it is time-consuming, and both the sophisticated equipment and well-trained personnel are required. Immunoassays and genetic diagnosis are expensive and limited to specificity and sensitivity. Aptamers are single-stranded deoxyribonucleic acid (ssDNA) and ribonucleic acid (RNA) oligonucleotide or peptide sequence generated in vitro based on the binding affinity of aptamer-target by a process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX). By taking several advantages over monoclonal antibodies and other conventional small-molecule therapeutics, such as high specificity and affinity, negligible batch-to-batch variation, flexible modification and production, thermal stability, low immunogenicity and lack of toxicity, aptamers are presently becoming promising novel diagnostic and therapeutic agents. This review describes the prospective application of aptamerbased laboratory diagnostic assays and therapeutics for pathogenic bacteria and toxins in bloodstream infections.

Acetate Kinase (AcK) is Essential for Microbial Growth and Betel-derived Compounds Potentially Target AcK, PhoP and MDR Proteins in M. tuberculosis, V. cholerae and Pathogenic E. coli: An in silico and in vitro Study

2019 ◽  
Vol 18 (31) ◽  
pp. 2731-2740 ◽  
Author(s):  
Sandeep Tiwari ◽  
Debmalya Barh ◽  
M. Imchen ◽  
Eswar Rao ◽  
Ranjith K. Kumavath ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
In Silico ◽  
Pathogenic Bacteria ◽  
In Vitro Study ◽  
Docking Studies ◽  
Acetate Kinase ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Novel Target

Background: Mycobacterium tuberculosis, Vibrio cholerae, and pathogenic Escherichia coli are global concerns for public health. The emergence of multi-drug resistant (MDR) strains of these pathogens is creating additional challenges in controlling infections caused by these deadly bacteria. Recently, we reported that Acetate kinase (AcK) could be a broad-spectrum novel target in several bacteria including these pathogens. Methods: Here, using in silico and in vitro approaches we show that (i) AcK is an essential protein in pathogenic bacteria; (ii) natural compounds Chlorogenic acid and Pinoresinol from Piper betel and Piperidine derivative compound 6-oxopiperidine-3-carboxylic acid inhibit the growth of pathogenic E. coli and M. tuberculosis by targeting AcK with equal or higher efficacy than the currently used antibiotics; (iii) molecular modeling and docking studies show interactions between inhibitors and AcK that correlate with the experimental results; (iv) these compounds are highly effective even on MDR strains of these pathogens; (v) further, the compounds may also target bacterial two-component system proteins that help bacteria in expressing the genes related to drug resistance and virulence; and (vi) finally, all the tested compounds are predicted to have drug-like properties. Results and Conclusion: Suggesting that, these Piper betel derived compounds may be further tested for developing a novel class of broad-spectrum drugs against various common and MDR pathogens.

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