UV-B辐射增强对陆地植物次生代谢的影响

平流层臭氧的减薄已导致地表中波紫外辐射(UV-B,280～320nm)增强,由于UV-B能被许多生物大分子如蛋白质和核酸吸收并引起分子构象的变化,因此可对植物的各方面产生影响。本文将近年来特别是近5年的UV-B辐射增强对植物次生代谢物影响的研究工作进行了综述。主要包括：UV-B辐射增强对植物紫外吸收物的影响和可能的机制;环境因子的复合作用对植物紫外吸收物的影响和可能的机制;UV-B辐射增强对次生代谢物影响的生态学意义。并对该领域未来的研究作了展望。     

A review of conditions affecting the radiolysis due to40K on nucleic acid bases and their derivatives adsorbed on clay minerals: Implications in prebiotic chemistry

This paper describes the possible effects of ionizing radiation arising from long-lived soluble radionuclides within clays, in particular⁴⁰K, at the epoch of the emergence of life on Earth. The free dispersion of soluble radionuclides constitutes an effectivein situ irradiation mechanism that might have acted upon adsorbed nucleic bases and their derivatives on clays, inducing chemical changes on these organic molecules. Several types of well documented reactions for radiolysis of nucleic acid bases and their derivatives are known, even at low doses (i.e., 0.1 Gy). For example, estimates with a dose rate calculated from⁴⁰K from deep sea clays at 3.8 Ga ago, indicates that over a period of 1000 years the amount of organic material transformated is 1.8 × 10^–7 moles/kg-clay.Although ionizing radiation may also induce synthetic reactions with prebiological interest, all in all these considerations indicate that nucleic acid bases and their derivatives adsorbed on clays were exposed for long periods to degradation conditions. Such situation promotes decomposition of organic molecules rather than protection of them and enhancement of further polymerization, as it has been usually taken for granted.     

Analysis of cellular components in immunocompetent organs treated with ionizing radiations]

The nucleic acid and protein content has been evaluated in lymphoid cells from the thymus and the Bursa of Fabricius of chicks exposed to different doses of radiation. A different response to radiation damage of the two cell populations is suggested by these analysis when compared with ultrastructural data.     

DNA conformation and dynamics

Nucleotide conformation and dynamics are important for the study of radiation damage to DNA at the atomic level. It is necessary to study not only normal oligonucleotide structure but also those containing modified bases which result from interaction with OH-radicals. There are now over 8000 atomic coordinate entries in the Brookhaven Protein Data Bank, of which over 900 relate to experimentally determined structures of nucleic acids and nucleic acid/protein complexes. We review some of these data which have led to the elucidation of novel DNA conformations, insight into DNA sequence specificity and knowledge of protein/DNA interactions. Further understanding of the conformation, stability and dynamics of nucleic acids has come from molecular modelling. We have used such techniques to study chemical modifications to bases such as alkylation of thymine and guanine and the effects of curvature in longer sequences. Recent improvements in this area include the inclusions of explicit counter-ions and solvent molecules, the use of Particle Mesh Ewald methods to incorporate the long-range electrostatic interactions and the use of longer time scale simulations. We have employed these methods to analyse the effects of incorporation of 8-oxodeoxyguanosine into duplex DNA. This lesion is a common result of radiation damage and is known to have important effects in mutagenesis, cancer and ageing. Received: 7 October 1998 / Accepted in revised form: 18 January 1999     

Control of development in plants and fungi by far-UV radiation

Far-UV (200–320 nm) radiation regulates development in plants and fungi. Some of these responses are controlled by a chromophore which absorbs strongly near 260 nm, possibly a nucleic acid. Other responses are controlled by a chromophore(s). with maximal in vivo sensitivity near 295 nm. In plants. far-UV induces genes in the phenylpropanoid pathway and the synthesis of phytoalexins and flavonoids. Far-UV also regulates growth rate. controls curvature and taxis, and stimulates sexual and asexual morphogenesis of fungi. Some of these developmental responses may prevent damage by far-UV radiation.     

Voltammetric determination of gamma radiation-induced DNA damage

Homopolydeoxyribonucleotides, poly[dGuo], poly[dAdo], poly[dThd], and poly[dCyd], and calf thymus single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) aqueous solutions previously exposed to gamma radiation doses between 2 and 35 Gy, were studied by differential pulse voltammetry using a glassy carbon electrode. The interpretation of the voltammetric data was also supported by the electrophoretic migration profile obtained for the same ssDNA and dsDNA gamma-irradiated samples by nondenaturing agarose gel electrophoresis. The generation of 8-oxo-7,8-dihydroguanine, 2,8-dihydroxyadenine, 5-formyluracil, base-free sites, and single- and double-stranded breaks in the gamma-irradiated DNA samples was detected voltammetrically, with the amount depending on the irradiation time. It was found that the current peaks obtained for 8-oxoguanine increase linearly with the radiation dose applied to the nucleic acid sample, and values between 8 and 446 8-oxo-7,8-dihydroguanine (8-oxoGua) per 10(6) guanines per Gy were obtained according to the nucleic acid sample. The results showed that voltammetry can be used for monitoring and simultaneously characterizing different kinds of DNA damage caused by gamma radiation exposure.     

Ultraviolet radiation, evolution and the π-electron system

Ultraviolet radiation is an important natural mutagen. Because of the energetic characteristics, the carbon compounds most susceptible to UV absorbance are those that contain π-electron systems. The π-electron configuration is most commonly represented in organic chemistry within aromatic ring structures. An analysis of a wide range of biochemically important processes shows that the susceptibility of this system lies at the heart of almost all UV radiation effects on life. Its disruption accounts for UV radiation-induced damage in nucleic acids, proteins and lipids. However, throughout the evolution of the biosphere, life has also used the UV absorbance of π-electron containing compounds to screen out UV radiation, turning their UV absorbance into a protection mechanism. Although UV radiation effects can be analyzed in terms of organism physiology, a more reductionist analysis shows the π-electron system to be the common chemical determinant in the evolution of UV radiation damage effects and protection strategies in organisms. It reveals an interesting evolutionary story.     

3种海洋赤潮微藻蛋白质和核酸合成对UV-B辐射增强的响应

应用同位素标志法, 研究了赤潮异弯藻 (Heterosigmaakashiwo) 、亚历山大藻 (Alexandriumtamarense ) 和中肋骨条藻 (Skeletonemacostatum) 核酸和蛋白质合成对紫外线B波段 (UV_B, 2 80~ 32 0nm) 辐射增强的响应变化。结果表明 :1) 按照由高到低的顺序, 3种海洋赤潮微藻对UV_B辐射增强的敏感性依次是赤潮异弯藻、亚历山大藻和中肋骨条藻。 2 ) UV_B辐射增强抑制赤潮异弯藻的生长和脱氧核糖核酸 (DNA) 的合成, 而低剂量的UV_B辐射对中肋骨条藻和亚历山大藻的生长与DNA的合成表现出刺激作用, 高剂量则表现出抑制作用。 3) 随着UV_B辐射的增强, 3种海洋赤潮微藻核糖核酸 (RNA) 和蛋白质的合成速度下降, 其中赤潮异弯藻合成速度的下降幅度明显大于中肋骨条藻和亚历山大藻, 表明赤潮异弯藻RNA和蛋白质的合成对UV_B辐射增强的敏感性高于中肋骨条藻和亚历山大藻。     

Effect of testosterone on nucleic acid synthesis in spermatogenic epithelial cells of irradiated animals

A study was made of changes in nucleic acid synthesis in proliferating germ cell populations of animals irradiated with doses of 2 and 4 Gy. The administration of exogenous testosterone was shown to stimulate DNA synthesis, inhibited by the effect of radiation, in spermatogonium populations of A1-4, intermediate, and B types, and in preleptotene primary spermatocytes; RNA synthesis was increased in the same populations of spermatogonia and pachytene primary spermatocytes. The obtained results indicate that exogenous testosterone exerts a beneficial effect on spermatogenesis damaged by ionizing radiation.     

Natural microbial UV radiation filters--mycosporine-like amino acids

Ozone depletion by anthropogenic gases has increased the atmospheric transmission of solar ultraviolet-B radiation (UV-B, 280-315 nm). There is a logical link between the natural defenses of terrestrial and marine organisms against UV radiation and the prevention of UV-induced damage to human skin. UV light degrades organic molecules such as proteins and nucleic acids, giving rise to structural changes that directly affect their biological function. These compounds offer the potential for development of novel UV blockers for human use. The biological role of mycosporine-like amino acids (MAAs) and scytonemin as a defense against solar radiation in organisms, together with their structure, synthesis, distribution, regulation and effectiveness, are reviewed in this article. This review points to the role of MAAs as a natural defense against UV radiation.     

Crystallographic studies of DNA and RNA

Our knowledge of nucleic acid structure grew rapidly over the past decade with the determination to high resolution of larger structures of great biological significance. Advances in sample preparation, crystallization techniques, cryocrystallography, access to synchrotron radiation, and crystallographic software continue to accelerate the structure determination of nucleic acids. Crystallographic studies of DNA and RNA molecules share many considerations that we outline here. The application of crystallography to RNA is illustrated with the structure determination of the CUG repeat that is linked to type I myotonic dystrophy.     

Selenium Derivatization of Nucleic Acids for X‐Ray Crystal‐Structure and Function Studies

It is estimated that over two thirds of all new crystal structures of proteins are determined via the protein selenium derivatization (selenomethionine (Se‐Met) strategy). This selenium derivatization strategy via MAD (multi‐wavelength anomalous dispersion) phasing has revolutionized protein X‐ray crystallography. Through our pioneer research, similarly, Se has also been successfully incorporated into nucleic acids to facilitate the X‐ray crystal‐structure and function studies of nucleic acids. Currently, Se has been stably introduced into nucleic acids by replacing nucleotide O‐atom at the positions 2′, 4′, 5′, and in nucleobases and non‐bridging phosphates. The Se derivatization of nucleic acids can be achieved through solid‐phase chemical synthesis and enzymatic methods, and the Se‐derivatized nucleic acids (SeNA) can be easily purified by HPLC, FPLC, and gel electrophoresis to obtain high purity. It has also been demonstrated that the Se derivatization of nucleic acids facilitates the phase determination via MAD phasing without significant perturbation. A growing number of structures of DNAs, RNAs, and protein–nucleic acid complexes have been determined by the Se derivatization and MAD phasing. Furthermore, it was observed that the Se derivatization can facilitate crystallization, especially when it is introduced to the 2′‐position. In addition, this novel derivatization strategy has many advantages over the conventional halogen derivatization, such as more choices of the modification sites via the atom‐specific substitution of the nucleotide O‐atom, better stability under X‐ray radiation, and structure isomorphism. Therefore, our Se‐derivatization strategy has great potentials to provide rational solutions for both phase determination and high‐quality crystal growth in nucleic‐acid crystallography. Moreover, the Se derivatization generates the nucleic acids with many new properties and creates a new paradigm of nucleic acids. This review summarizes the recent developments of the atomic site‐specific Se derivatization of nucleic acids for structure determination and function study. Several applications of this Se‐derivatization strategy in nucleic acid and protein research are also described in this review.     

Interaction of a natural modifier of lethal effect of radiation with HeLa cells

Polypeptide 14C-aspartyl glutamate, a chemical analogue of a natural modified of a lethal effect of radiation, has been synthesized. The modifier was shown to react readily with nucleic acids and proteins of non-irradiated cells: the reaction was considerably enhanced when the modifier was administered simultaneously with radiation of cells. As is known from the literature, a molar concentration of the incorporated polypeptide is considerably lower than the intracellular molar concentration of an active radiosensitizer, BUdr, which produces an effect on mammalian cells comparable with that of polypeptide.     

Laser cross-linking of nucleic acids to proteins. Methodology and first applications to the phage T4 DNA replication system

Single-pulse (approximately 8 ns) ultraviolet laser excitation of protein-nucleic acid complexes can result in efficient and rapid covalent cross-linking of proteins to nucleic acids. The reaction produces no nucleic acid-nucleic acid or protein-protein cross-links, and no nucleic acid degradation. The efficiency of cross-linking is dependent on the wavelength of the exciting radiation, on the nucleotide composition of the nucleic acid, and on the total photon flux. The yield of cross-links/laser pulse is largest between 245 and 280 nm; cross-links are obtained with far UV photons (200-240 nm) as well, but in this range appreciable protein degradation is also observed. The method has been calibrated using the phage T4-coded gene 32 (single-stranded DNA-binding) protein interaction with oligonucleotides, for which binding constants have been measured previously by standard physical chemical methods (Kowalczykowski, S. C., Lonberg, N., Newport, J. W., and von Hippel, P. H. (1981) J. Mol. Biol. 145, 75-104). Photoactivation occurs primarily through the nucleotide residues of DNA and RNA at excitation wavelengths greater than 245 nm, with reaction through thymidine being greatly favored. The nucleotide residues may be ranked in order of decreasing photoreactivity as: dT much greater than dC greater than rU greater than rC, dA, dG. Cross-linking appears to be a single-photon process and occurs through single nucleotide (dT) residues; pyrimidine dimer formation is not involved. Preliminary studies of the individual proteins of the five-protein T4 DNA replication complex show that gene 43 protein (polymerase), gene 32 protein, and gene 44 and 45 (polymerase accessory) proteins all make contact with DNA, and can be cross-linked to it, whereas gene 62 (polymerase accessory) protein cannot. A survey of other nucleic acid-binding proteins has shown that E. coli RNA polymerase, DNA polymerase I, and rho protein can all be cross-linked to various nucleic acids by the laser technique. The potential uses of this procedure in probing protein-nucleic acid interactions are discussed.     

Electronic spectra, excited state structures and interactions of nucleic acid bases and base assemblies: a review

A comprehensive review of recent theoretical and experimental advances in the singlet electronic transitions, excited state structures and dynamics of nucleic acid bases (NABs) and base assemblies are presented. It is well known that NABs absorb ultraviolet radiation, but the absorbed energy is efficiently dissipated in the form of ultrafast internal conversion processes believed to occur in the subpicosecond time scale and, therefore, enabling NABs highly photostable. It is not known how much evolutionary role was played in evolving these molecules and the ultimate selection by nature as genetic materials, but it is well accepted that survival-of-fittest prevails. Recently, significant efforts have been continuously paid to understand the mechanism of electronic excitation deactivation, but universally acceptable mechanism is still elusive. However, recent investigations reveal that electronic excited state geometries of DNA bases are usually nonplanar and this structural nonplanarity may facilitate nonradiative deactivation. Investigation of excited state structures is challenging and, therefore, it is not surprising that despite the impressive theoretical and computational advances, this research area is still hampered by the methodological and computational limitations. Further, stacking has significant influence on the emission properties of molecules. The 2-aminopurine, a fluorescent adenine derivative frequently used in studying DNA dynamics, shows significant attenuations in fluorescence quantum yield when incorporated in the DNA. Theoretical and computational bottlenecks limit a thorough theoretical understanding of effect of stacking interactions on the excited state dynamics of NABs. Despite these limitations the investigations of excited state properties are progressing in the right direction and our better understanding of excited state structure and dynamics of NABs and nucleic acids may help to design preventive strategy for radiation induced illness and photostable materials.     

Radiation Inhibition of Amino Acid Uptake by Escherichia coli

The inhibition of macromolecular synthesis in Escherichia coli by ionizing radiation has been investigated. The survival of the ability to incorporate arginine, leucine, isoleucine, histidine, uracil, and glucose after various doses of gamma radiation, deuteron and alpha particle bombardment has been measured. All amino acids are incorporated by processes which show the same radiation sensitivity. The sensitivity of uracil corresponds to a volume which is roughly spherical, of radius about 160A, whereas the amino acids possess sensitive regions which are long and thin in character. The uptake of glucose is concerned with a smaller, roughly spherical unit. The possible identification of the radiation-sensitive targets with cellular constituents is discussed. The long thin character observed for amino acids suggests that the sensitive region affected by radiation is an unfolded form of a ribosome, or alternatively a long nucleic acid molecule. For uracil the sensitive region fits with a 70S ribosome, while for glucose a smaller particle would fit the data.     

Photoprotection of DNA (in vitro) by acyclothymidine dinucleosides

Excessive exposure to sunlight is primarily implicated in ultraviolet (UV) induced skin cancers worldwide. Direct absorption of UV radiation by DNA leads to the formation of cyclobutane pyrimidine dimers (CPDs) resulting in DNA damage. The molecular mechanisms involved in the mutagenicity of CPDs are well established. Photoprotection of the skin from the detrimental effects of UV is essential in preventing skin damage. A variety of formulations, which essentially contain UV filters have been used as photoprotective agents of the skin. These comprise aromatic and inorganic molecules, whose mechanism of action involves either absorption, reflection, or scattering of UV radiation. However, the downstream photoproducts of some of these molecules have undesirable characteristics which compromise their utility. A biomimetic approach involving structural analogs of nucleic acids can help overcome these limitations. Herein, we show the photoprotective action of acyclothymidine dinucleosides on both plasmid and cellular DNA.     

Effects of Ionizing Radiation on Bacterial DNA-membrane Complexes

THE model proposed by Alper¹ for lethal radiation damage to cells is based on inferential evidence that there are two important sites of damage by ionizing radiation. At one site, damage referred to as type “N” is associated with a low oxygen enhancement ratio (OER) and is probably to nucleic acid, while at the other site, type “O” damage is associated with a considerably higher oxygen enhancement value and is to a non-nucleic acid target. The model demands that the two values of OER are respectively less and greater than that observed for the overall lethal effect. More recently² Alper reviewed further inferential evidence³ that cell membranes are the site of type O damage, though there may be subsequent interaction with the lesions following energy deposition in DNA⁴.