112 research outputs found
Identification and evaluation of the role of the manganese efflux protein in Deinococcus radiodurans
<p>Abstract</p> <p>Background</p> <p><it>Deinococcus radiodurans </it>accumulates high levels of manganese ions, and this is believed to be correlated with the radiation resistance ability of this microorganism. However, the maintenance of manganese ion homeostasis in <it>D. radiodurans </it>remains to be investigated.</p> <p>Results</p> <p>In this study, we identified the manganese efflux protein (MntE) in <it>D. radiodurans</it>. The null mutant of <it>mntE </it>was more sensitive than the wild-type strain to manganese ions, and the growth of the <it>mntE </it>mutant was delayed in manganese-supplemented media. Furthermore, there was a substantial increase in the <it>in vivo </it>concentration of manganese ions. Consistent with these characteristics, the <it>mntE </it>mutant was more resistant to H<sub>2</sub>O<sub>2</sub>, ultraviolet rays, and Ξ³-radiation. The intracellular protein oxidation (carbonylation) level of the mutant strain was remarkably lower than that of the wild-type strain.</p> <p>Conclusions</p> <p>Our results indicated that <it>dr1236 </it>is indeed a <it>mntE </it>homologue and is indispensable for maintaining manganese homeostasis in <it>D. radiodurans</it>. The data also provide additional evidence for the involvement of intracellular manganese ions in the radiation resistance of <it>D. radiodurans</it>.</p
Identification and evaluation of the role of the manganese efflux protein in Deinococcus radiodurans
<p>Abstract</p> <p>Background</p> <p><it>Deinococcus radiodurans </it>accumulates high levels of manganese ions, and this is believed to be correlated with the radiation resistance ability of this microorganism. However, the maintenance of manganese ion homeostasis in <it>D. radiodurans </it>remains to be investigated.</p> <p>Results</p> <p>In this study, we identified the manganese efflux protein (MntE) in <it>D. radiodurans</it>. The null mutant of <it>mntE </it>was more sensitive than the wild-type strain to manganese ions, and the growth of the <it>mntE </it>mutant was delayed in manganese-supplemented media. Furthermore, there was a substantial increase in the <it>in vivo </it>concentration of manganese ions. Consistent with these characteristics, the <it>mntE </it>mutant was more resistant to H<sub>2</sub>O<sub>2</sub>, ultraviolet rays, and Ξ³-radiation. The intracellular protein oxidation (carbonylation) level of the mutant strain was remarkably lower than that of the wild-type strain.</p> <p>Conclusions</p> <p>Our results indicated that <it>dr1236 </it>is indeed a <it>mntE </it>homologue and is indispensable for maintaining manganese homeostasis in <it>D. radiodurans</it>. The data also provide additional evidence for the involvement of intracellular manganese ions in the radiation resistance of <it>D. radiodurans</it>.</p
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Distinct lipid membrane interaction and uptake of differentially charged nanoplastics in bacteria
Background
Nanoplastics have been recently found widely distributed in our natural environment where ubiquitously bacteria are major participants in various material cycles. Understanding how nanoplastics interact with bacterial cell membrane is critical to grasp their uptake processes as well as to analyze their associated risks in ecosystems and human microflora. However, little is known about the detailed interaction of differentially charged nanoplastics with bacteria. The present work experimentally and theoretically demonstrated that nanoplastics enter into bacteria depending on the surface charges and cell envelope structural features, and proved the shielding role of membrane lipids against nanoplastics.
Results
Positively charged polystyrene nanoplastics (PS-NH2, 80Β nm) can efficiently translocate across cell membranes, while negatively charged PS (PS-COOH) and neutral PS show almost no or much less efficacy in translocation. Molecular dynamics simulations revealed that the PS-NH2 displayed more favourable electrostatic interactions with bacterial membranes and was subjected to internalisation through membrane penetration. The positively charged nanoplastics destroy cell envelope of Gram-positive B. subtilis by forming membrane pore, while enter into the Gram-negative E. coli with a relatively intact envelope. The accumulated positively charged nanoplastics conveyed more cell stress by inducing a higher level of reactive oxygen species (ROS). However, the subsequently released membrane lipid-coated nanoplastics were nearly nontoxic to cells, and like wise, stealthy bacteria wrapped up with artifical lipid layers became less sensitive to the positively charged nanoplastics, thereby illustrating that the membrane lipid can shield the strong interaction between the positively charged nanoplastics and cells.
Conclusions
Our findings elucidated the molecular mechanism of nanoplasticsβ interaction and accumulation within bacteria, and implied the shielding and internalization effect of membrane lipid on toxic nanoplastics could promote bacteria for potential plastic bioremediation.
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Regulation of MntH by a Dual Mn(II)- and Fe(II)-Dependent Transcriptional Repressor (DR2539) in Deinococcus radiodurans
The high intracellular Mn/Fe ratio observed within the bacteria Deinococcus radiodurans may contribute to its remarkable resistance to environmental stresses. We isolated DR2539, a novel regulator of intracellular Mn/Fe homeostasis in D. radiodurans. Electrophoretic gel mobility shift assays (EMSAs) revealed that DR2539 binds specifically to the promoter of the manganese acquisition transporter (MntH) gene, and that DR0865, the only Fur homologue in D. radiodurans, cannot bind to the promoter of mntH, but it can bind to the promoter of another manganese acquisition transporter, MntABC. Ξ²-galactosidase expression analysis indicated that DR2539 acts as a manganese- and iron-dependent transcriptional repressor. Further sequence alignment analysis revealed that DR2539 has evolved some special characteristics. Site-directed mutagenesis suggested that His98 plays an important role in the activities of DR2539, and further protein-DNA binding activity assays showed that the activity of H98Y mutants decreased dramatically relative to wild type DR2539. Our study suggests that D. radiodurans has evolved a very efficient manganese regulation mechanism that involves its high intracellular Mn/Fe ratio and permits resistance to extreme conditions
A Novel OxyR Sensor and Regulator of Hydrogen Peroxide Stress with One Cysteine Residue in Deinococcus radiodurans
In bacteria, OxyR is a peroxide sensor and transcription regulator, which can sense the presence of reactive oxygen species and induce antioxidant system. When the cells are exposed to H2O2, OxyR protein is activated via the formation of a disulfide bond between the two conserved cysteine residues (C199 and C208). In Deinococcus radiodurans, a previously unreported special characteristic of DrOxyR (DR0615) is found with only one conserved cysteine. dr0615 gene mutant is hypersensitive to H2O2, but only a little to ionizing radiation. Site-directed mutagenesis and subsequent in vivo functional analyses revealed that the conserved cysteine (C210) is necessary for sensing H2O2, but its mutation did not alter the binding characteristics of OxyR on DNA. Under oxidant stress, DrOxyR is oxidized to sulfenic acid form, which can be reduced by reducing reagents. In addition, quantitative real-time PCR and global transcription profile results showed that OxyR is not only a transcriptional activator (e.g., katE, drb0125), but also a transcriptional repressor (e.g., dps, mntH). Because OxyR regulates Mn and Fe ion transporter genes, Mn/Fe ion ratio is changed in dr0615 mutant, suggesting that the genes involved in Mn/Fe ion homeostasis, and the genes involved in antioxidant mechanism are highly cooperative under extremely oxidant stress. In conclusion, these findings expand the OxyR family, which could be divided into two classes: typical 2-Cys OxyR and 1-Cys OxyR
The Special Relationship Between Nepenthes and Tree Frogs
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