Dataset Information

Enhancement of DNaseI Salt Tolerance by Mimicking the Domain Structure of DNase from an Extremely Halotolerant Bacterium Thioalkalivibrio sp. K90mix.

ABSTRACT: In our previous work we showed that DNaseI-like protein from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix retained its activity at salt concentrations as high as 4 M NaCl and the key factor allowing this was the C-terminal DNA-binding domain, which comprised two HhH (helix-hairpin-helix) motifs. The further investigations revealed that this domain originated from proteins related to bacterial competence ComEA/ComE proteins. It is likely that in the course of evolution the DNA-binding domain from these proteins was fused to a metallo-?-lactamase superfamily domain. Very likely such domain organization having proteins subsequently "donated" the DNA-binding domain to bacterial DNases. In this study we have mimicked this evolutionary step by fusing bovine DNaseI and DNA-binding domains. We have created two fusions: one harboring the DNA-binding domain of DNaseI-like protein from Thioalkalivibrio sp. K90mix and the second one harboring the DNA-binding domain of bacterial competence protein ComEA from Bacillus subtilis. Both domains enhanced salt tolerance of DNaseI, albeit to different extent. Molecular modeling revealed the essential differences between their interaction with DNA shedding some light on the differences in salt tolerance. In this study we have enhanced salt tolerance of bovine DNaseI; thus, we successfully mimicked the Nature's evolutionary engineering that created the extremely halotolerant bacterial DNase. We have demonstrated that the newly engineered DNaseI variants can be successfully used in applications where activity of the wild type bovine DNaseI is impeded by buffers used.

SUBMITTER: Alzbutas G

PROVIDER: S-EPMC4777378 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Publications

Enhancement of DNaseI Salt Tolerance by Mimicking the Domain Structure of DNase from an Extremely Halotolerant Bacterium Thioalkalivibrio sp. K90mix.

Alzbutas Gediminas G Kaniusaite Milda M Lagunavicius Arunas A

PloS one 20160303 3

In our previous work we showed that DNaseI-like protein from an extremely halotolerant bacterium Thioalkalivibrio sp. K90mix retained its activity at salt concentrations as high as 4 M NaCl and the key factor allowing this was the C-terminal DNA-binding domain, which comprised two HhH (helix-hairpin-helix) motifs. The further investigations revealed that this domain originated from proteins related to bacterial competence ComEA/ComE proteins. It is likely that in the course of evolution the DNA- ...[more]

PMID: 26939122

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Project description:A characteristic trait of plants living in harsh environments is their association with fungal endophytes, which enable them to survive under extreme stress. Abiotic stress resistance in agro-ecosystems, particularly in arid and semi-arid regions, can be increased by inoculating these fungal endophytes on plants other than their original hosts. The present study is therefore focused on the possible role of three halotolerant endophytic fungi, i.e., Periconia macrospinosa, Neocamarosporium goegapense, and N. chichastianum, isolated from roots of salt lake plants growing in the central desert of Iran, in alleviating the adverse effects of salinity and drought stresses on barley under greenhouse conditions. To perform this experiment, a randomized block design was applied with three factors: fungi (four levels including three halotolerant endophytic species and control), salinity (three levels including 8, 12, and 16 dS/m), and drought (four levels including 100, 80, 60, 40 percent field capacity). All plants were measured for growth characteristics, chlorophyll concentration, proline content, and antioxidant enzyme activities. A three-way analysis of variance indicated that all three fungal endophytes, to varying extents, induced the barley plants’ resistance to salinity and drought, and their combined effects. Additionally, we found that fungal endophytes were more effective when the barley plants were subjected to higher levels of salinity and drought. Under the stress of salinity and drought, a strong relationship between inoculation of fungal endophytes and enhancement of biomass, shoot length, chlorophyll concentration, proline content, and activity of catalase, peroxidase, and superoxide dismutase was indicated. We discussed that increased root growth, proline content, and antioxidant enzyme activity are the main physiological and biochemical mechanisms causing stress resistance in barley plants inoculated with endophytes. Our research findings illustrate that fungal endophytes have a substantial potential for increasing abiotic stress tolerance in barley plants, which can be applied in agricultural ecosystems.

Dataset Information

Enhancement of DNaseI Salt Tolerance by Mimicking the Domain Structure of DNase from an Extremely Halotolerant Bacterium Thioalkalivibrio sp. K90mix.

Publications

Enhancement of DNaseI Salt Tolerance by Mimicking the Domain Structure of DNase from an Extremely Halotolerant Bacterium Thioalkalivibrio sp. K90mix.

Similar Datasets

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