Investigation of the zinc uptake system of the human fungal pathogen Candida parapsilosis

Candida parapsilosis is the second or third most commonly isolated Candida species from blood cultures and is frequently associated with infections in neonatal intensive care units. Candida species have several virulence factors enabling them to adapt to host environmental conditions and cause infections. These factors include adhesion, biofilm formation, and secretion of hydrolytic enzymes, such as acidic proteinases and lipases. Candida species also obtain heavy metal ions from their environment, such as zinc. Zinc is a cofactor of several proteins and a vital element in cellular mechanisms of the fungi. On the one hand, the host niche represents a zinc-limited environment, that indirectly inhibits microbial growth. In order to survive in such an environment, these pathogens have evolved a zinc transport system that allows them to access bound zinc ions during infection. On the other hand, high zinc ion concentration within the host can also be toxic to microbes e.g. in the phagosomes of Mycobacterium tuberculosis infected macrophages. In case of C. albicans, zinc acquisition processes are intensively studied, but we lack information of the zinc uptake, transfer and homeostasis mechanisms in C. parapsilosis. Here, predicted potential zinc transporters in C. parapsilosis using in silico analyses, generated homozygous knock out mutants and performed their phenotypical characterization by exposing them to various types of stressors and zinc limiting conditions. Furthermore, we analyzed their virulence traits by examining kinetics of fungal cell uptake by macrophages, their killing efficiency and also investigated zinc ion levels in the phagolysosome during in vitro infections

Biochemical Analysis Based on Zinc Uptake of Chickpea (Cicer arietinum L.) Varieties Infected by Meloidogyne incognita

The significant constraints in Chickpea (Cicer arietinum L.) production hampers a bit more than 14% global yield loss due to plant-parasitic nematodes. Root-knot nematode (Meloidogyne sp.) is an endoparasite and a significant species affecting the chickpea plant. So, the chemical basis of management is more cost-effective, and pest resurgence building is enhanced in the pathogen. So, ecological-based nematode management is requisite, which also is got hampered due to breeding for resistance against such plant-parasitic nematodes. This was the primary reason to conduct this experiment to enhance resistance in the chickpea plants based on Zinc uptake by using bioagent, Pseudomonas fluorescens alone or in combination. where Different treatments including nematode, bacterium, and chemicals were used sustaining the enhancement of disease resistance in chickpea cultivars, RSG 974, GG 5, GNG 2144. Zinc content of chickpea variety GNG 2144 was found the highest in treatment, when only bacterium (P. fluorescens) was inoculated, i.e., 3.14 mg/100g of root followed by GG 5, i.e., 2.79 mg/100g of root and RSG 974 was, i.e., 2.35 mg/100g of root respectively in a descending order. Application of P. fluorescence combined or alone gradually increased the Zn concentration in roots of chickpea plants compared to healthy check followed by chemical treated plants.

Zinc uptake and distribution in ivy (Hedera helix L.) leaves

Detached leaves of ivy (Hedera helix L.) were used as a model for the study of zinc uptake and transport in vascular plants. By the uptake via the surface of fully immersed leaves in 25 % Hoagland nutrient media (HM) spiked with 65ZnCl2 (50 μmol/dm3 ZnCl2), concentration in leaves 4.98 μg Zn/g (dry wt.), i. e. 2.6 μg Zn/dm2 leaf area after 7d exposition were obtained. By the uptake via immersed stalks of not immersed (transpiring) leaves concentrations up to 370 μg Zn/g (dry wt.) were obtained. When Zn enters into detached leaves via the surface of immersed leaf blades, zinc is uniformly distributed in leaf blades and leaf stalks. When zinc enters detached leaves via immersed stalks of non-immersed transpiring leaves, only small part of zinc is transported to leaf blades and the prevailing part remains in leaf stalks. Stalks act as a trap, able to prevent other leaf tissues against inhibitory effects of high Zn concentrations. Mineral nutrient salts in solutions mobilize Zn trapped in leaf stalks and facilitate Zn transport by transpiration stream to leaf blades, what means that Zn in stalks is bound in ion-exchageable forms.

Slc39a2-Mediated Zinc Homeostasis Modulates Innate Immune Signaling in Phenylephrine-Induced Cardiomyocyte Hypertrophy

Zinc dyshomeostasis has been involved in the pathogenesis of cardiac hypertrophy; however, the dynamic regulation of intracellular zinc and its downstream signaling in cardiac hypertrophy remain largely unknown. Using Zincpyr1 staining, we found a significant decrease of intracellular Zinc concentration in phenylephrine (PE)-induced hypertrophy of neonatal rat ventricular myocytes (NRVMs). We then screened SLC39 family members responsible for zinc uptake and identified Slc39a2 as the only one altered by PE treatment. Slc39a2 knockdown in NRVMs reduced the intracellular Zinc level, and exacerbated the hypertrophic responses to PE treatment. In contrast, adenovirus-mediated Slc39a2 overexpression enhanced zinc uptake and suppressed PE-induced Nppb expression. RNA sequencing analysis showed a pro-hypertrophic transcriptome reprogramming after Slc39a2 knockdown. Interestingly, the innate immune signaling pathways, including NOD signaling, TOLL-like receptor, NFκB, and IRFs, were remarkably enriched in the Slc39a2-regulated genes. Slc39a2 deficiency enhanced the phosphorylation of P65 NFκB and STAT3, and reduced the expression of IκBα. Finally, the expression of IRF7 was significantly increased by Slc39a2 knockdown, which was in turn suppressed by IRF7 knockdown. Our data demonstrate that zinc homeostasis mediated by a Slc39a2/IRF7 regulatory circuit contributes to the alteration of innate immune signaling in cardiomyocyte hypertrophy.

Strategies for Zinc Uptake in Pseudomonas aeruginosa at the Host–Pathogen Interface

As a structural, catalytic, and signaling component, zinc is necessary for the growth and development of plants, animals, and microorganisms. Zinc is also essential for the growth of pathogenic microorganisms and is involved in their metabolism as well as the regulation of various virulence factors. Additionally, zinc is necessary for infection and colonization of pathogenic microorganisms in the host. Upon infection in healthy organisms, the host sequesters zinc both intracellularly and extracellularly to enhance the immune response and prevent the proliferation and infection of the pathogen. Intracellularly, the host manipulates zinc levels through Zrt/Irt-like protein (ZIP)/ZnT family proteins and various zinc storage proteins. Extracellularly, members of the S100 protein family, such as calgranulin C, sequester zinc to inhibit microbial growth. In the face of these nutritional limitations, bacteria rely on an efficient zinc transport system to maintain zinc supplementation for proliferation and disruption of the host defense system to establish infection. Here, we summarize the strategies for zinc uptake in conditional pathogenic Pseudomonas aeruginosa, including known zinc uptake systems (ZnuABC, HmtA, and ZrmABCD) and the zinc uptake regulator (Zur). In addition, other potential zinc uptake pathways were analyzed. This review systematically summarizes the process of zinc uptake by P. aeruginosa to provide guidance for the development of new drug targets.

The zinc transporter ZnuABC is critical for the virulence of Chromobacterium violaceum and contributes to diverse zinc-dependent physiological processes

Chromobacterium violaceum is a ubiquitous environmental bacterium that causes sporadic life-threatening infections in humans. How C. violaceum acquires zinc to colonize environmental and host niches is unknown. In this work, we demonstrated that C. violaceum employs the zinc uptake system ZnuABC to overcome zinc limitation in the host, ensuring the zinc supply for several physiological demands. Our data indicated that the C. violaceum ZnuABC transporter is encoded in a zur -CV_RS15045-CV_RS15040- znuCBA operon. This operon was repressed by the zinc uptake regulator Zur and derepressed in the presence of the host protein calprotectin (CP) and the synthetic metal chelator EDTA. A Δ znuCBA mutant strain showed impaired growth under these zinc-chelated conditions. Moreover, the deletion of znuCBA provoked a reduction in violacein production, swimming motility, biofilm formation, and bacterial competition. Remarkably, the Δ znuCBA mutant strain was highly attenuated for virulence in an in vivo mouse infection model and showed a low capacity to colonize the liver, grow in the presence of CP, and resist neutrophil killing. Overall, our findings demonstrate that ZnuABC is essential for C. violaceum virulence, contributing to subvert the zinc-based host nutritional immunity.