The impact of exposure to excess zinc on the transcriptome of Streptococcus mutans
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PROVIDER: PRJNA702540 | ENA |
REPOSITORIES: ENA
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Project description:Zinc is a micronutrientient that serves as an important co-factor necessary for enzymatic function. In the host environment, access to metals can be highy restricted. As excess metals can be toxic to the bacterial cell, the host immune system can utilize zinc intoxication as a defense mechanism against bacterial infection. Here, we wanted to identify the transcriptomic response when S. mutans UA159 is exposed to an excess of zinc.
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Project description:Zinc is an essential divalent trace metal for living cells. Intracellular zinc homeostasis is critical to the pathogenicity and virulence of bacteria, however the strategies of zinc homeostasis maintain in Streptococcus mutans, an important causative pathogen of dental caries, is unknown. In this study, we examined the function of an ABC transporter AdcCBA and its transcription factor AdcR in S. mutans. Using RNA-Seq analysis, LacZ-reporter studies and gel mobility shift assay, we showed that a MarR family transcription factor AdcR directly regulates the activity of promoters of adcRCB and adcA in response to the concentration of zinc in their environmental niches. Deletion of adcR increases the sensitivity of S. mutans to excessive zinc supply. Taken together, our finding suggests that Adc regulon, consists of a zinc uptake transporter AdcCBA and a zinc-responsive repressor AdcR, plays a prominent role in the maintenance of intracellular zinc homeostasis of S. mutans.
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Project description:Antimicrobial resistance (AMR) is a global health crisis that is predicted to worsen. While improper antibiotic usage is an established driver, less is known on the impacts of metal supplements. Here, we probe the impact of zinc (Zn) on AMR. In conflict settings where diarrhea disease cases are high, Zn, which is associated with weapons of war, is given as a supplement for diarrhea treatment prior to antibiotics such as ciprofloxacin. In this study, we find that E. coli’s exposure order to zinc impacts resistance development, with increasing pre-exposure leading to accelerated ciprofloxacin resistance, while combined exposure of zinc with ciprofloxacin delays ciprofloxacin resistance. We did not find evidence that zinc pre-exposure leads to genetic changes or changes in antibiotic tolerance, though the lag phase and doubling time of E. coli was increased, suggesting gene expression may be changed. While the zinc phenotype was no longer observed if ciprofloxacin exposure did not occur right after zinc pre-exposure, the resulting elevated MIC was more stable. These results are important as they highlight the need to reexamine the clinical role of zinc in treating diarrheal diseases and assess if changes in resistance development observed in vitro are also observed in vivo.
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Project description:A common reference model was used to design the microarray experiment, in which cDNA from T. caerulescens roots exposed to 100 uM (sufficient) zinc was used as the common reference. Every slide was always hybridized with the common reference sample and with a sample from one of the treatments (Arabidopsis or T. caerulescens exposed to deficient, sufficient or excess zinc). The common reference was labeled with the fluorescent dye Cyanine 3 (Cy3) and the treatment samples were labeled with Cyanine 5 (Cy5). As a quality control step we performed a dye-swap hybridization for one sample (from T. caerulescens roots exposed to sufficient zinc).
2006-10-10 | E-MEXP-877 | biostudies-arrayexpress
Project description:Transcriptional Profiling of Streptococcus mutans UA159 Grown in Continuous Culture using TV Media Supplemented With 10 mM vs 100 mM Glucose. The genetic and phenotypic responses of Streptococcus mutans, an organism known to be strongly associated with the development of dental caries, to changes in carbohydrate availability were investigated. S. mutans UA159 or a derivative of UA159 lacking ManL, which is the EIIAB component (EIIABMan) of a mannose/glucose permease of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) and a dominant effector of catabolite repression, were grown in continuous culture to steady-state in conditions of excess (100 mM) or limiting (10 mM) glucose. Microarrays using RNA from S. mutans UA159 revealed that 174 genes were differentially expressed in response to changes in carbohydrate availability (P < 0.001). Glucose-limited cells possessed higher PTS activity, could acidify the environment more rapidly and to a greater extent, and produced more ManL protein than cultures grown with excess glucose. Loss of ManL adversely affected carbohydrate transport and acid tolerance. Comp arison of the HPr protein in S. mutans UA159 and the manL deletion strain indicated that the differences in behaviors of the strains were not due to major differences in HPr pools or HPr phosphorylation status. Therefore, carbohydrate availability alone can dramatically influence the expression of physiologic and biochemical pathways that contribute directly to the virulence of S. mutans, and ManL has a profound influence on this behavior. Two-condition experiment, growth in 10 mM vs 100 mM glucose. Biological replicates: 3 per condition, independently grown and harvested. One replicate per array
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Project description:Deficiency of the micronutrient zinc is a widespread condition in agricultural soils, generating a negative impact on crop quality and yield. Nevertheless, there is insufficient knowledge on the regulatory and molecular mechanisms underlying the plant response to inadequate zinc nutrition. Hydroponically-grown Arabidopsis thaliana plants (WT Col-0 and the T-DNA insertion double mutant bzip19 bzip23) were subjected to zinc deficiency, sufficiency and excess. The particular role of the bZIP19 and bZIP23 transcription factors was investigated via the bzip19 bzip23 double mutant.
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