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 H₂O₂, ultraviolet rays, and γ-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 H₂O₂, ultraviolet rays, and γ-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

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. β-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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A Novel Classification Optimization Approach Integrating Class Adaptive MRF and Fuzzy Local Information for High Spatial Resolution Multispectral Imagery

This paper develops a novel classification optimization approach integrating class adaptive Markov Random Field (MRF) and fuzzy local information (CAMRF-FLI) for high spatial resolution multispectral imagery (HSRMI). Firstly, the raw classification results, including initial fuzzy memberships and class labels of every pixel, are achieved by a pixel-wise classification method for a given image. Secondly, the class adaptive MRF-based data energy function is developed to integrate class spatial dependency information. Thirdly, a novel spatial energy function integrating fuzzy local information is constructed. Finally, based on the total of data and spatial energies, the raw classification map is regularized by a global minimization of the energy function using its iterated conditional modes (ICM). The effectiveness of CAMRF-FLI is performed by two data sets. The results indicate it can refine the classification map in homogeneous areas, meanwhile, reduce most of the edge blurring artifact, and improve the classification accuracy compared with some conventional approaches