Physiologic determinants of radiation resistance in Deinococcus radiodurans

Immense volumes of radioactive wastes, which were generated during nuclear weapons production, were disposed of directly in the ground during the Cold War, a period when national security priorities often surmounted concerns over the environment. The bacterium Deinococcus radiodurans is the most radiation-resistant organism known and is currently being engineered for remediation of the toxic metal and organic components of these environmental wastes. Understanding the biotic potential of D. radiodurans and its global physiological integrity in nutritionally restricted radioactive environments is important in development of this organism for in situ bioremediation. We have previously shown that D. radiodurans can grow on rich medium in the presence of continuous radiation (6,000 rads/h) without lethality. In this study we developed a chemically defined minimal medium that can be used to analyze growth of this organism in the presence and in the absence of continuous radiation; whereas cell growth was not affected in the absence of radiation, cells did not grow and were killed in the presence of continuous radiation. Under nutrient-limiting conditions, DNA repair was found to be limited by the metabolic capabilities of D. radiodurans and not by any nutritionally induced defect in genetic repair. The results of our growth studies and analysis of the complete D. radiodurans genomic sequence support the hypothesis that there are several defects in D. radiodurans global metabolic regulation that limit carbon, nitrogen, and DNA metabolism. We identified key nutritional constituents that restore growth of D. radiodurans in nutritionally limiting radioactive environments.     

Fungal diversity in the Antarctic active layer

Taxonomic diversity of fungi in the samples of the active layer of Antarctica was investigated using conventional microbiological techniques and metagenomic analysis of total DNA extracted from environmental samples. The list of Antarctic microscopic fungi was expanded, including detection of the species representing a portion of the fungal complex which is nonculturable or sterile on conventional nutrient media.     

Enzymes of Direct,Excision, and Mismatch DNA Repair in Pro- and Eukaryotes and Their Biological Role

DNA repair is of paramount importance to protect the integrity of the cell genome. Over the last decade, DNA repair enzymes have been intensely studied owing to their role in mutagenesis, carcinogenesis, and aging. This led to discoveries of novel enzymes, protein factors as well as their intermolecular complexes involved in the repair process. Alternative pathways of cell response to the same type of damage were identified. Data on the tertiary structure of a number of enzymes and mechanisms of their molecular interaction were obtained. This paper is an overview of latest advances in the research of cell response to DNA damage via direct, base and nucleotide excision, and mismatch DNA repair.     

Three-Domain Antibodies against the Tumor Necrosis Factor: Investigation of Their Biological Activity In Vitro

Russian Journal of Bioorganic Chemistry - The aim of this work was to evaluate novel three-domain antibodies consisting of two domains specific for human tumor necrosis factor (hTNF), and of the...     

Reclassification of Methylarcula marina Doronina et al. 2000 as Paracoccus methylarcula nom. nov.

Microbiology - Phylogenetic analysis of the 16S rRNA gene sequences showed that the type species of the genus Methylarcula, Methylarculamarina h1T was closely related to Paracoccus saliphilus DSM...     

Protein oxidation implicated as the primary determinant of bacterial radioresistance

In the hierarchy of cellular targets damaged by ionizing radiation (IR), classical models of radiation toxicity place DNA at the top. Yet, many prokaryotes are killed by doses of IR that cause little DNA damage. Here we have probed the nature of Mn-facilitated IR resistance in Deinococcus radiodurans, which together with other extremely IR-resistant bacteria have high intracellular Mn/Fe concentration ratios compared to IR-sensitive bacteria. For in vitro and in vivo irradiation, we demonstrate a mechanistic link between Mn(II) ions and protection of proteins from oxidative modifications that introduce carbonyl groups. Conditions that inhibited Mn accumulation or Mn redox cycling rendered D. radiodurans radiation sensitive and highly susceptible to protein oxidation. X-ray fluorescence microprobe analysis showed that Mn is globally distributed in D. radiodurans, but Fe is sequestered in a region between dividing cells. For a group of phylogenetically diverse IR-resistant and IR-sensitive wild-type bacteria, our findings support the idea that the degree of resistance is determined by the level of oxidative protein damage caused during irradiation. We present the case that protein, rather than DNA, is the principal target of the biological action of IR in sensitive bacteria, and extreme resistance in Mn-accumulating bacteria is based on protein protection.     

A Summary of Recent Results in the Study of Taimyr Amber

Paleontological Journal - Two families, two tribes, 30 genera and 57 insect species are described as new from Taimyr amber, as well as the first new plant genus and species to be described from...     

Variations of the effect of insulin on neutrophil respiratory burst. The role of tyrosine kinases and phosphatases

The priming effect of insulin on the fMLP-induced respiratory burst of mouse neutrophils as well as the involvement of tyrosine protein kinases and phosphatases in this process have been studied. Peritoneal evoked neutrophils of NMRI strain mice were incubated with 0.01-100 nM insulin for 1-60 min at 22, 30, or 37°C and activated by 0.1-50 M N-formyl-methionyl-leucyl-phenylalanine (fMLP). The production of reactive oxygen species (ROS) by neutrophils was monitored by luminol-dependent chemiluminescence. We found that ¹²⁵I-labeled insulin binding by mouse neutrophils occurred with saturation and high affinity. Insulin itself did not change the basal level of the ROS production but could modulate fMLP-induced respiratory burst. The effect of insulin depended on temperature and duration of pretreatment of the neutrophils with insulin and the concentration combination of the insulin and fMLP. The tyrosine kinase inhibitor tyrphostin 51 decreased the fMLP-induced respiratory burst significantly. Insulin did not change the fMLP response of neutrophils pretreated with tyrphostin. However, the effect of tyrphostin on the response to 50 M fMLP was considerably decreased in neutrophils treated with insulin. There was no such effect during activation by 5 M fMLP, for which the priming effect of insulin was not observed. Insulin did not increase the fMLP-induced respiratory burst in neutrophils treated with the protein phosphatase inhibitors orthovanadate and pyrophosphate. If the inhibitors were added after insulin, the combined effect was nearly additive. It is possible that priming by insulin of the fMLP-induced respiratory burst is triggered by tyrosine phosphorylation, realized with its participation, and involves the signaling pathways initiated by tyrosine phosphorylation but subsequently is not dependent on the latter. The role of protein phosphatases in priming by insulin is of little importance. The data indirectly confirm the idea that priming of the neutrophil respiratory burst is a result of crosstalk of signaling pathways of the insulin and fMLP receptors with the participation of tyrosine phosphorylation.