Functional identification of catalytic metal ion binding sites within RNA

The viability of living systems depends inextricably on enzymes that catalyze phosphoryl transfer reactions. For many enzymes in this class, including several ribozymes, divalent metal ions serve as obligate cofactors. Understanding how metal ions mediate catalysis requires elucidation of metal ion interactions with both the enzyme and the substrate(s). In the Tetrahymena group I intron, previous work using atomic mutagenesis and quantitative analysis of metal ion rescue behavior identified three metal ions (MA, MB, and MC) that make five interactions with the ribozyme substrates in the reaction's transition state. Here, we combine substrate atomic mutagenesis with site-specific phosphorothioate substitutions in the ribozyme backbone to develop a powerful, general strategy for defining the ligands of catalytic metal ions within RNA. In applying this strategy to the Tetrahymena group I intron, we have identified the pro-SP phosphoryl oxygen at nucleotide C262 as a ribozyme ligand for MC. Our findings establish a direct connection between the ribozyme core and the functionally defined model of the chemical transition state, thereby extending the known set of transition-state interactions and providing information critical for the application of the recent group I intron crystallographic structures to the understanding of catalysis.     

Induction of active forms of oxygen by biotic elicitors in tomato cell cultures

The elicitor-induced generation of two oxygen species in tomato cell culture as well as their involvement into hypersensitive reaction was investigated. Generation of superoxide O2.- was measured by a lucigenin-related chemiluminescence. Accumulation of hydrogen peroxide H2O2 was measured by a fluorescent probe pyranin. Xylanase and chitosan were used as biotic elicitors with different mode of action. It was found that both O2.- and H2O2 had been accumulated in elicitor-treated tomato cells. The results obtained show that reactive oxidants are important signal transduction elements for activation of hypersensitive response in tomato cells.     

Singlet oxygen scavenging activity of tocopherol and plastochromanol in Arabidopsis thaliana: relevance to photooxidative stress

In the present study, singlet oxygen (¹O₂) scavenging activity of tocopherol and plastochromanol was examined in tocopherol cyclase‐deficient mutant (vte1) of Arabidopsis thaliana lacking both tocopherol and plastochromanol. It is demonstrated here that suppression of tocopherol and plastochromanol synthesis in chloroplasts isolated from vte1 Arabidopsis plants enhanced ¹O₂ formation under high light illumination as monitored by electron paramagnetic resonance spin‐trapping spectroscopy. The exposure of vte1 Arabidopsis plants to high light resulted in the formation of secondary lipid peroxidation product malondialdehyde as determined by high‐pressure liquid chromatography. Furthermore, it is shown here that the imaging of ultra‐weak photon emission known to reflect oxidation of lipids was unambiguously higher in vte1 Arabidopsis plants. Our results indicate that tocopherol and plastochromanol act as efficient ¹O₂ scavengers and protect effectively lipids against photooxidative damage in Arabidopsis plants.     

Integral mechanisms of regulation of oxygen delivery and consumption in human body

With new algorithms of pattern recognition three types of regulation of oxygen delivery and consumption were studied: at normal regulation, hypo- and hyperfunction of heart. It is shown that a surplus systemic blood flow results in increase of arterio-venous shunt flow. Insufficient blood flow results in increase of arterio-venous gradient of blood oxygen content due to the increase of gradient of oxygen content on both sides of capillary wall and to activating of vasomotor function of microcirculatory arterial bed. Quantitative estimations of shunt and capillary blood flow are obtained.     

The Influence of quick-to-court Gene on Some Aspects of Mating Behavior in Drosophila melanogaster

Russian Journal of Genetics - quick-to-court (qtc) is a recently discovered gene involved in the regulation of the mating behavior of Drosophila melanogaster males. Previously, we obtained flies...     

The effect of phosphorylation on oxidative and CoA- and NAD+-independent turnover of pyruvate by pyruvate dehydrogenase complex in the brain

It was shown that in the presence of ATP and Mg2+ the phosphorylation of the partially purified pyruvate dehydrogenase complex and the enzyme in isolated brain mitochondria inhibited the oxidative activity of the pyruvate dehydrogenase complex. The phosphorylation did no affect essentially the nonoxidative decarboxylation of pyruvate to form CO2 and acetaldehyde. In native mitochondria from the bovine brain the nonoxidative activity of the pyruvate dehydrogenase complex reached about 10% as compared to the oxidative activity of enzyme.