On a possible mechanism of the effect of microwave radiation on biological macromolecules

A model that describes the dissociation of a hydrogen bond in water clusters when irradiated by an electromagnetic field in the microwave range is proposed. The model is also applicable for the case of the rupture of the covalent bond of the water molecule in a cluster. If the energy absorption occurs at the interface of water and polymer clusters (e.g., DNA and chitosan), degradation of the polymer chain is possible.     

The effect of heat and radiation on the initiation and elongation processes of DNA synthesis

The pH step alkaline elution and alkaline sucrose gradient techniques were utilized to evaluate alterations in DNA replication (initiation and elongation) induced by heat and low dose X-irradiation is synchronized Chinese hamster ovary cells. The initiation and elongation process of DNA synthesis were radioresistant at the G1/S boundary (4 hours after mitosis) while in mid S phase (9 hours after mitosis) DNA initiation and elongation were sensitive to X-irradiation. The initiation and elongation processes of DNA synthesis which were radiation resistant at the G1/S boundary could be inhibited by a hyperthermia treatment (43 degrees C for 1 hour beginning at 4 hours after mitosis). The impairment of initiation in the heated cells was maintained through late S phase while that of elongation was reversible as judged by full recovery at 15 hours after mitosis. These data suggest that the known synergistic lethality of heat and radiation may be mediated by an impairment of initiation of DNA synthesis.     

On the mechanism of premeiotic DNA synthesis in the yeast Saccharomyces cerevisiae

Premeiotic DNA synthesis in synchronously sporulating cultures of the yeast, Saccharomyces cerevisiae, was analysed by sedimentation in alkaline sucrose gradients and by DNA-fibre autoradiography. The gradient profiles of cells pulse-labelled for varying times were essentially identical with those obtained with mitotic cultures, revealing a close resemblance between the meiotic and mitotic replication mechanisms. This was supported by the finding that exposure of meiotic cells to a specific concentration of hydroxyurea led to the accumulation of completed, but unjoined replicons, just as it does in mitotic cells. The results of DNA-fibre autoradiography confirmed that replicons in meiotic cells are the same size (20–180 Kb, averaging around 90 Kb) as in mitotic cells, and assuming replication is bi-directional, replication forks must move at round the same rate as in mitosis, i.e. about 0.7 μm/min.     

DNA合成的忠实性机制

DNA的忠实性合成对于基因组稳定和物种延续至关重要,否则可能会产生严重的后果。DNA合成具有极高的忠实性,这主要基于3个步骤:(1)基于氢键、碱基对构象或其他因素的核苷酸选择;(2)基于3′→5′外切酶活性的校对,方式有顺式校对和反式校对,可以去除错误掺入的核苷酸;(3)基于错配修复、切除修复、同源重组修复和跨损伤DNA合成的修复过程,可以纠正逃过校对的错误核苷酸。由于DNA聚合酶不仅可以作为抗病毒或抗癌药物的靶标,而且其忠实性还与抗药性或药物副作用有关,所以深入研究DNA合成的忠实性具有非常重要的意义。文章主要论述了DNA合成的忠实性机制,并对DNA聚合酶的应用前景做了展望。     

The effect of gamma radiation on DNA methylation

The effect of 60Co gamma radiation on DNA methylation was studied in four cultured cell lines. In all cases a dose-dependent decrease in 5-methylcytosine was observed at 24, 48, and 72 h postexposure to 0.5-10 Gy. Nuclear DNA methyltransferase activity decreased while cytoplasmic activity increased in irradiated (10 Gy) V79A03 cells as compared to controls. No DNA demethylase activity was detected in the nuclei of control or irradiated V79A03 cells. Additionally, gamma radiation resulted in the differentiation of C-1300 N1E-115 cells, a mouse neuroblastoma line, in a dose- and time-dependent manner. These results are consistent with the hypothesis that (1) genes may be turned on following radiation via a mechanism involving hypomethylation of cytosine and (2) radiation-induced hypomethylation results from decreased intranuclear levels of DNA methyltransferase.     

Differential effect of aphidicolin on adenovirus DNA synthesis and cellular DNA synthesis.

There is strong evidence for a participation of DNA polymerase gamma in the replication of adenovirus (Ad) DNA. To study a possible additional role of DNA polymerase alpha we measured the effect of aphidicolin on viral DNA replication. In intact cells, aphidicolin inhibits Ad DNA synthesis weakly. The drug concentration required for 50% inhibition of Ad DNA replication was 300-400 fold higher than for a similar effect on cellular DNA synthesis. Such a differential inhibition was also observed in AGMK cells doubly infected with SV40 and the simian adenovirus SA7. No evidence was found for modification of aphidicolin in infected cells or for a change in aphidicolin sensitivity of DNA polymerase alpha after infection. The extent of inhibition of purified DNA polymerase alpha was dependent upon the dCTP concentration. The same situation was observed when DNA synthesis was studied in isolated nuclei from uninfected cells. However, in nuclei from Ad infected cells no effect of dCTP on aphidicolin sensitivity was found. These results were taken as evidence that DNA polymerase alpha does not participate in the replication of adenovirus DNA.     

On the mechanism of cytogenetic effect of electromagnetic radiation: a role of oxidation homeostasis

It was established in the experiments on rats that the changes in free radical oxidation under the influence of non-ionizing radiation had a wavy character. It was revealed that the changes in oxidation homeostasis preceded development of cytogenetic effects and could be their reason.     

Structure of human DNA polymerase iota and the mechanism of DNA synthesis

Cellular DNA polymerases belong to several families and carry out different functions. Highly accurate replicative DNA polymerases play the major role in cell genome replication. A number of new specialized DNA polymerases were discovered at the turn of XX–XXI centuries and have been intensively studied during the last decade. Due to the special structure of the active site, these enzymes efficiently perform synthesis on damaged DNA but are characterized by low fidelity. Human DNA polymerase iota (Pol ι) belongs to the Y-family of specialized DNA polymerases and is one of the most error-prone enzymes involved in DNA synthesis. In contrast to other DNA polymerases, Pol ι is able to use noncanonical Hoogsteen interactions for nucleotide base pairing. This allows it to incorporate nucleotides opposite various lesions in the DNA template that impair Watson-Crick interactions. Based on the data of X-ray structural analysis of Pol ι in complexes with various DNA templates and dNTP substrates, we consider the structural peculiarities of the Pol ι active site and discuss possible mechanisms that ensure the unique behavior of the enzyme on damaged and undamaged DNA.     

Extent of excision repair before DNA synthesis determines the mutagenic but not the lethal effect of UV radiation

Excision repair-proficient diploid fibroblasts from normal persons (NF) and repair-deficient cells from a xeroderma pigmentosum patient (XP12BE, group A) were grown to confluence and allowed to enter the G₀ state. Autoradiography studies of cells released from G₀ after 72 h and replated at lower densities (3?9 × 10³ cells/cm²) in fresh medium containing 15% fetal bovine serum showed that semiconservative DNA synthesis (S phase) began ～24 h after the replating. To determine whether the time available for DNA excision repair between ultraviolet irradiation (254 nm) and the onset of DNA synthesis was critical in determining the cytotoxic and/or mutagenic effect of UV in human fibroblasts, we released cultures of NF or XP12BE cells from G₀, allowed them to reattach at lower densities, irradiated them in early G₁ (～18 h prior to the onset of S) or just prior to S phase, and assayed the frequency of mutations to 6-thioguanine resistance and the survival of colony-forming ability. The XP12BE cells, which are virtually incapable of excising UV-induced DNA lesions, showed approximately the same frequency of mutations and survival regardless of the time of UV irradiation. In NF cells, the slope of the dose response for mutations induced in cells irradiated just prior to S was about 7-fold steeper than that of cells irradiated 18 h earlier. However, the two sets of NF cells showed no significant difference in survival. Neither were there significant differences in the survival of NF cells released from G₀, plated at cloning densities and irradiated as soon as they had attached and flattened out (～20 h prior to S) or 4, 8, 12, 16, 20 or 24 h later. We conclude that the frequency of mutations induced by UV is dependent upon the number of unexcised lesions remaining at the time of semi-conservative DNA replication. However, the amount of time available for excision of potentially cytotoxic lesions is not determined primarily by the period between irradiation and the onset of S phase.