Environmental RNA interference in animals

Animals interact with their environments all the time, and must react to all kinds of factors present in the environments. Environmen- tal RNA interference refers to the phenomenon that exogenous RNA molecules can enter cells of certain animal species and inter- fere with gene expression of these animals. These exogenous RNAs may be molecules in animal food, or may be present in the living environment from other sources. Molecular pathways for the cell entry of environmental RNAs and further for their func- tionality in gene interference have started to be revealed in the animal model Caenorhabditis elegans. Here we highlight some known examples of environmental RNA interference （RNAi） in animals and introduce the molecular mechanisms underneath.     

有效制备核糖核酸的方法研究

核糖核酸的制备和分析受多种因素的影响．因此，在特定条件下制备高纯度和完整性好的ＲＮＡ的方法研究至关重要．本文以大白鼠肝为材料，研究ＲＮＡ的制备方法，建立了有效的ＲＮＡ制备方法．用紫外分光光度计和变性电泳分析所制备的ＲＮＡ，发现其纯度Ａ２６０Ａ２８０＝１．７～２．０，２８ＳｒＲＮＡ与１８ＳｒＲＮＡ的带宽比为２１．本方法具有节约节时，重复性好，产量较高，无ＤＮＡ污染等优点．这为ｃＤＮＡ合成，基因表达和调控等研究奠定了基础．     

Evidence for hydrogen-deuterium exchange in viral particles

Heavy water (D2O) could enhance thermostability of some viruses. However, the underlying mechanism is not clear. Here we report the development of a matrix-aided gas-isotope-ratio mass spectrometry method that allows direct determination of deuterium/hydrogen (D/H) ratio in D2O-treated Japanese encephalitis virus (JEV), hepatitis A virus (HAV) and RNA from D2O-treated HAV. The D/H ratiowas expressed as δDSMOW. Our experiments showed that δDSMOW values increased significantly in D2O-treated viral samples compared to normal controls, and increment in δDSMOW of D2O, treated viral samples was in a fine linear relationship with increment in amount of samples loaded in BSA matrix. Our experiments also indicated that increased δDSMOW of D2O-treated virus correlated well with its enhanced thermostability. The results suggested that hydrogen-deuterium exchange occurred in viral particles and its RNA structure as a result of D2O-treatment. Furthermore,such exchange could cause changes in viral phenotype, such as enhanced thermostability.     

计算最大堆迭的RNA二级结构预测算法

RNA二级结构预测用于蛋白质功能分析，在生物信息学研究中具有重要意义．提出了一个时间复杂度为O（n^2）的基于Greedy算法思想的算法．基于“堆迭结构相对稳定”的RNA分子结构特征，算法思想为计算具有最多堆迭的RNA二级结构．用VC＋＋编程实现了该算法，采用PseudoBase的RNA分子片段进行了计算实验，结果表明该算法具有良好的准确度．该算法可预测RNA分子的嵌套二级结构和伪结点一级结构．     

肌肉抑制素基因与RNA干扰的应用前景

本文综述了肌肉抑制素基因的研究进展,以及RNA干扰技术的功能.展望了RNA干扰技术在肌肉抑制素基因中的应用前景.     

丝瓜核糖体失活蛋白的研究进展综述

丝瓜核糖体失活蛋白是一类具多种生物学功能和活性的蛋白,本文在阐述了该类蛋白的理化性质的同时，综述了该类蛋白的酶学机制、活性位点和用做免疫毒素等方面的研究进展。     

MicroRNA特征与功能

通过分析总结现代分子生物学国际前沿microRNA(miRNA)领域的研究文献，整理出miRNA研究的基本脉络和走向。miRNA是一类长度～22nt的非编码小分子RNA，在包括线虫、果绳、家鼠、人体以及拟南芥等生物中普遍存在；它在调节基因转录与表达，调控生物体正常发育等生理过程中扮演重要角色。从比较的角度出发，揭示了miRNA与小干扰RNA在其代谢与功能方面共用某些途径，相互交叉与替代，可能同属一个更广范围的小分子RNA介导的生理调控机制。miRNA的研究可能对新一代基因药物的开发具有深远意义。     

The multi-KH protein vigilin associates with free and membrane-bound ribosomes

The-multi-KH domain protein vigilin has been identified by ex vivo experiments as both a tRNA- and/or mRNA-binding protein. We show here that in vitro under conditions previously shown to allow tRNA binding, recombinant vigilin also binds to selected mRNA species and ribosomal RNA. An in vivo link of vigilin to mRNA and rRNA was elucidated by several approaches. (i) Coexpression/costimulation of vigilin was found with many other proteins independently of whether their mRNA was translated on free or membrane-bound ribosomes. (ii) A close codistribution of vigilin with free ribosomes was seen in the cytoplasm while nucleoli were a major organelle of vigilin accumulation in the nucleus. (iii) Furthermore, free and membrane-bound ribosomes can be enriched for vigilin which suggests that this binding does not depend on the class of mRNA translated. Therefore, we suggest that vigilin does not distinguish between free or membrane-bound ribosomes but is generally necessary for the localization of mRNAs to actively translating ribosomes.Received 20 June 2003; received after revision 25 July 2003; accepted 29 July 2003     

The contribution of noncatalytic phosphate-binding subsites to the mechanism of bovine pancreatic ribonuclease A

The enzymatic catalysis of polymeric substrates such as proteins, polysaccharides or nucleic acids requires precise alignment between the enzyme and the substrate regions flanking the region occupying the active site. In the case of ribonucleases, enzyme-substrate binding may be directed by electrostatic interactions between the phosphate groups of the RNA molecule and basic amino acid residues on the enzyme. Specific interactions between the nitrogenated bases and particular amino acids in the active site or adjacent positions may also take place. The substrate-binding subsites of ribonuclease A have been characterized by structural and kinetic studies. In addition to the active site (p₁ ), the role of other noncatalytic phosphate-binding subsites in the correct alignment of the polymeric substrate has been proposed. p₂ and p₀ have been described as phosphate-binding subsites that bind the phosphate group adjacent to the 3′ side and 5′ side, respectively, of the phosphate in the active site. In both cases, basic amino acids (Lys-7 and Arg-10 in p₂ , and Lys-66 in p₀ ) are involved in binding. However, these binding sites play different roles in the catalytic process of ribonuclease A. The electrostatic interactions in p₂ are important both in catalysis and in the endonuclease activity of the enzyme, whilst the p₀ electrostatic interaction contributes only to binding of the RNA.