U.V. enhanced reactivation of U.V.-and gamma-irradiated adenovirus in normal human fibroblasts

An enhanced reactivation (UVER) of U.V.-irradiated as well as of gamma-irradiated human adenovirus type 2 (Ad 2) was detected following infection of normal human fibroblasts which had been pre-irradiated with U.V. light. U.V.-irradiated or non-irradiated fibroblasts were infected with either non-irradiated or irradiated Ad 2, and at 48 hours after infection cells were examined for the presence of viral structural antigens (Vag) using immunofluorescent staining. Results obtained using 5 different normal fibroblast strains showed that irradiation of host monolayers with 10J/m2 immediately prior to infection gave a U.V. enhanced reactivation (UVER) factor +/- standard error equal to 3 . 1 +/- 1 . 2 for virus U.V.-irradiated with 1 . 2 x 10(3) J/m2, and 2 . 1 +/- 0 . 5 for virus gamma-irradiated with 2 x 10(4) Gy. For a fixed survival of about 5 . 9 x 10(-2) for irradiated virus, the efficiency of UVER for gamma-irradiated virus was about 0 . 18, slightly less than the value of about 0 . 24 obtained for U.V.-irradiated virus. The results of time course experiments indicated that while U.V.-irradiation of normal host monolayers prior to infection gave rise to an increased rate of Vag formation for infection by unirradiated Ad 2, U.V.-irradiation of the cells increased the proportion of cells able to repair U.V.-damaged virus as well as allowing an earlier onset and/or increased rate of synthesis of Vag from a U.V.-damaged template. Similar experiments involving gamma-ray enhanced reactivation (gamma-RER) of irradiated Ad 2 indicated that gamma-RER and UVER may operate, in part at least, by different mechanisms in normal human cells.     

Initiation of Bacillus subtilis Ribonucleic Acid Polymerase on Deoxyribonucleic Acid from Bacteriophages 2C, φ29, T4, and Lambda

Ribonucleic acid (RNA) synthesis primed by bacteriophage T4 or lambda deoxyribonucleic acid (DNA) with Bacillus subtilis RNA polymerase is severely inhibited by high ionic strength. In contrast, RNA synthesis on B. subtilis bacteriophage 2C, SPO1, or phi29 DNA is only moderately affected under similar conditions. The basis of this inhibition lies in the inability of the enzyme to initiate RNA chains with adenosine triphosphate or guanosine triphosphate (ATP, GTP). Binding to templates and the rate of catalysis in high salt after initiation do not seem to be affected. Incorporation of gamma-(32)P-ATP and GTP under a variety of conditions suggests that the specificity of B. subtilis RNA polymerase is different from that of the Escherichia coli enzyme and that it recognizes few promoters on T4 and lambda DNA. Although B. subtilis RNA polymerase initiates RNA chains primarily with ATP or GTP, initiations with pyrimidines can occur on DNA molecules in which hydroxymethyluracil replaces thymine. RNA synthesis on denatured DNA does not seem to be inhibited by high ionic strength, and on native T4 or lambda DNA the inhibition of initiation at constant ionic strength is inversely but not linearly proportional to the ionic radii of cations used to stabilize bihelical DNA to denaturation.     

Lack of enhanced reactivation of simian virus 40 irradiated with van de Graaff electrons in U.V.-irradiated CV-1 monkey cells

Simian virus 40 (SV40) irradiated with U.V. or van de Graaff electrons was assayed on CV-1 monkey cells irradiated with U.V. before virus infection. U.V.-irradiated cells enhanced the survival of U.V.-irradiated virus, while little or no enhancement was observed for electron-irradiated virus assayed on U.V.-irradiated cells. It is suggested that the U.V.-irradiated cells are able to increase the repair of U.V.-damaged viral DNA, but not of electron-damaged DNA.     

色氨酸操纵子调控机理详析

色氨酸操纵子是最早被研究的细菌合成代谢调控、基因表达调控的模型之一。其中阻遏蛋白对转录起始的抑制作用、色氨酸作为辅阻遏物的作用以及通过定点突变揭示的弱化作用的分子机制已基本被阐明。此外,色氨酸操纵子RNA结合弱化蛋白、NusA、NusG、TrpY等调节蛋白对细菌色氨酸操纵子弱化作用的调节机制也在近年来得到进一步揭示。特别是在枯草芽孢杆菌中,色氨酸操纵子主要依赖于转录衰减机制调控,包括由色氨酸激活的色氨酸操纵子RNA结合弱化蛋白与新生转录产物结合形成内部终止子,导致5′非翻译区（5′UTR）转录终止。NusA、NusG通过刺激RNA聚合酶在5′UTR的U107和U144位点暂停,释放出RNA聚合酶,最终造成转录终止。不同的是,在U144位点NusA参与的转录弱化机制依赖其发夹结构,且NusA与RNA聚合酶作用促进了RNA结合弱化蛋白与新生转录产物的结合,使转录终止。而NusG是通过与非模板DNA链中的一段富含T碱基序列和RNA聚合酶同时互作,阻止了RNA聚合酶向下游移动,从而引起RNA聚合酶高效停滞。但在细菌操纵子中,绝大多数调节因子参与的弱化机制最终依赖于ρ因子,从而导致多达一半的转录终止事件发生。近年来,随着学科的发展,越来越多关于色氨酸操纵子调节机制新概念被挖掘报道,这也使人类对色氨酸操纵子的表达调控机制的认知愈加详尽。