Enzymes involved in DNA ligation and end-healing in the radioresistant bacterium Deinococcus radiodurans

BACKGROUND: Enzymes involved in DNA metabolic events of the highly radioresistant bacterium Deinococcus radiodurans are currently examined to understand the mechanisms that protect and repair the Deinococcus radiodurans genome after extremely high doses of gamma-irradiation. Although several Deinococcus radiodurans DNA repair enzymes have been characterised, no biochemical data is available for DNA ligation and DNA endhealing enzymes of Deinococcus radiodurans so far. DNA ligases are necessary to seal broken DNA backbones during replication, repair and recombination. In addition, ionizing radiation frequently leaves DNA strand-breaks that are not feasible for ligation and thus require end-healing by a 5'-polynucleotide kinase or a 3'-phosphatase. We expect that DNA ligases and end-processing enzymes play an important role in Deinococcus radiodurans DNA strand-break repair. RESULTS: In this report, we describe the cloning and expression of a Deinococcus radiodurans DNA ligase in Escherichia coli. This enzyme efficiently catalyses DNA ligation in the presence of Mn(II) and NAD+ as cofactors and lysine 128 was found to be essential for its activity. We have also analysed a predicted second DNA ligase from Deinococcus radiodurans that is part of a putative DNA repair operon and shows sequence similarity to known ATP-dependent DNA ligases. We show that this enzyme possesses an adenylyltransferase activity using ATP, but is not functional as a DNA ligase by itself. Furthermore, we identified a 5'-polynucleotide kinase similar to human polynucleotide kinase that probably prepares DNA termini for subsequent ligation. CONCLUSION: Deinococcus radiodurans contains a standard bacterial DNA ligase that uses NAD+ as a cofactor. Its enzymatic properties are similar to E. coli DNA ligase except for its preference for Mn(II) as a metal cofactor. The function of a putative second DNA ligase remains unclear, but its adenylyltransferase activity classifies it as a member of the nucleotidyltransferase family. Characterization of another protein from the same operon revealed a 5'-polynucleotide kinase with a possible role in DNA strand-break repair

Experimental Research of the Diffraction and Vavilov-Cherenkov Radiation Generation in a Teflon Target

Geometry of Vavilov-Cherekov (VChR) radiation when an electron moves close to a dielectric target is in analogy to diffraction radiation (DR) geometry. In this case we may expect DR generation from the upstream face of the target besides that VChR. The joint observation of these booth types of radiation is very interesting from the pseudo-photon viewpoint, which is applicable for relativistic electrons. Unexpected results obtained in our experiment insist on reflection about nature both DR and VChR. The experiment was performed on the relativistic electron beam of the microtron of Tomsk Polytechnic University.Comment: This article will be published in Journal of Physic

Electromagnetic field features at interaction of relativistic electron with matter

The features of electromagnetic field of relativistic electrons passing through a hole in an absorbing screen as a function of the distance from the screen in the range of radiation formation length were investigated for the transversal and longitudinal field components. The analysis of the obtained results allows approving the existence of a semi-bare electron with a particularly deprived Coulomb field, which turns into the stable state of the usual electron at the distance of radiation formation length

Experimental test of the shadowing effect in Smith-Purcell radiation

The observation of a shadowing effect of a relativistic electron Coulomb field for the Smith-Purcell radiation generation is presented in this paper. For this purpose the surface current from the closest surface of grating element to the electron beam was measured for a downstream one shadowed by upstream element. The experimental results showed that shadowing effect for Smith-Purcell radiation depends on grating geometry

Evolution of the Velocity Ellipsoids in the Thin Disk of the Galaxy and the Radial Migration of Stars

Data from the revised Geneva--Copenhagen catalog are used to study the influence of radial migration of stars on the age dependences of parameters of the velocity ellipsoids for nearby stars in the thin disk of the Galaxy, assuming that the mean radii of the stellar orbits remain constant. It is demonstrated that precisely the radial migration of stars, together with the negative metallicity gradient in the thin disk,are responsible for the observed negative correlation between the metallicities and angular momenta of nearby stars, while the angular momenta of stars that were born at the same Galactocentric distances do not depend on either age or metallicity. (abridged)Comment: Astronomy Reports, Vol. 86 No. 9, P.1117-1126 (2009

Modern approaches to artificial gene synthesis: aspects of oligonucleotide synthesis, enzymatic assembly, sequence verification and error correction

Synthetic biology is a rapidly developing field aimed at engineering of biological systems with predictable properties. Synthetic biology accumulates the achievements of modern biological sciences, programming and computational model­ing as well as engineering technologies for creation of biologi­cal objects with user-defined properties. Evolution of synthetic biology has been marked by a number of technological developments in each of the mentioned fields. Thus, significant reduction in cost of DNA sequencing has provided an easy access to large amounts of data on the genetic sequences of various organisms, and decreased the price of the DNA sequence synthesis, which, analogous to Moore’s law, resulted in an opportunity to create a lot of potential genes without the time – consuming and labor – intensive traditional methods of molecular biology. Development of system biology has allowed forming a deeper understanding of the functions and relationship of natural biological models, as well as of the computational models describing processes at the cell and system levels. Combination of these factors has created an op­portunity for conscious changes of natural biological systems. In this review the modern approaches to oligonucleotide gene assembly synthesis are discussed, including such aspects as protocols for gene assembly, sequence verification, error cor­rection and further applications of synthesized genes

On the training of specialists in the creation and maintenance of software for processing and presentation of geospatial data

Показана актуальність відкриття підготовки в вузах Білорусі програмістів геодезичного профілю.=The relevance of the opening of training in the universities of Belarus for geodetic programmers is shown