小核仁RNAs及其衍生RNAs的研究进展

小核仁RNAs(small nucleolarRNAs,snoRNAs)是一类发现较早且位于核仁内的小非编码RNAs,在核糖体RNAs(ribosomalRNAs,rRNAs)、信使RNAs(messengerRNAs,mRNAs)、小核RNAs(small nuclearRNAs,snRNAs)的成熟及修饰中均发挥重要作用。snoRNAs的功能及其作用途径一直以来均是学术界的研究热点。目前,snoRNAs对rRNAs的化学修饰作用已得到广泛认可。另外,有研究表明snoRNAs与一些遗传疾病以及肿瘤性疾病的发生发展存在密切关联。近年来研究发现,部分snoRNAs经切割可生成更小的、有功能的RNAs,即小核仁RNAs衍生RNAs(snoRNAs derivedRNAs,sdRNAs),这些sdRNAs中部分具有微小RNAs(microRNAs,miRNAs)的特征,可发挥类似miRNA的作用,这一发现极大地拓展了snoRNAs的作用机制方式。结合国内外研究现状,在总结snoRNAs的结构和基本功能的基础上对sdRNAs与miRNAs之间的相关性进行了综述,以期为后续的相关研究提供参考。     

Separation of α- and β-Globin Messenger RNAs

THE 10S RNA fraction of reticulocytes from various species contains the haemoglobin messenger RNA^1–4. When this 10S RNA fraction is added to a cell-free system derived from reticulocytes or Krebs II ascites cells, it directs the synthesis of α and β chains of haemoglobin^5–8. The α and β messenger RNA molecules contained in this fraction, however, have not yet been separated and identified. When reticulocyte. RNA of mouse is subjected to electrophoresis on 6% polyacrylamide gels, the 10S fraction contains two major bands and three minor bands⁹, suggesting that the major lOS RNA bands contain the messenger RNAs for the α- and β-globin chains.     

Emerging novel functions of RNAs, and binary phenotype?

Loss-of-function technology has been one of the most popular knockout tools for the study of gene function in cell and developmental biology. This technology employs two basic approaches for elimination of the protein of interest. The morpholino antisense oligonucleotides approach relies on inhibiting translation of the given protein without degrading the cognate mRNA. The antisense deoxynucleotides and siRNA approach acts via removal of the mRNA template, which then prevents protein translation. In the latest approach, as well as in these genetic knockout approaches that eliminate or alter the level of mRNA transcribed from the gene of interest, the assumption is and always has been that the only relevant function of mRNA is to make a protein, and, thus, the effect of removing mRNA equals the effect of removing its protein function. However, the most recent studies of different biological systems point to completely novel and unexpected functions of the subpopulation of localized RNAs and suggest that, at least in some cases, the normal cell or embryo phenotype is in fact binary i.e. depends not only on the function of the protein but also on the autonomous function of its mRNA.     

Compartmentation of uridine 5′-triphosphate occurs during synthesis of cytoplasmic and mitochondrial ribosomal RNAs of cultured tomato cells but not during synthesis of cytoplasmic ribosomal and transfer RNAs

Compartmentation of uridine 5-triphosphate (UTP) was studied during the nucleolar synthesis of cytoplasmic ribosomal RNA (cyt-rRNA) and the synthesis of cytoplasmic transfer RNA (cyt-tRNA) in the nuclear matrix as well as the synthesis of mitochondrial ribosomal RNA (mt-rRNA) in tomato (Lycopersicon esculentum Mill. cv. Lukullus) cell-suspension culture using the approach of Wiegers et al. (Eur. J. Biochem. 64, 535–540, 1976). Before measurements were made, it was ensured that: (i) there was steady-state labeling of all RNAs studied as well as UTP; (ii) there was stability of cyt-tRNA and cyt-rRNA; (iii) there was no label randomization through degradation of [³H]uridine; (iv) there were significant differences in the specific radioactivity of UTP, the final immediate precursor of RNA, after supplying the cells with two different exogenous [³H]uridine concentrations.By comparing the steady-state specific radioactivity of UTP with that of cyt-tRNA and cyt-18S rRNA during constant [³H]uridine supply, we found that the three molecules had equal specific radioactivities which, however, differed significantly from that of the mt-rRNA. With a 20-fold higher uridine concentration, i.e. a 20-fold lower specific radioactivity of exogenous [³H]uridine, the specific radioactivity of cyt-rRNA, cyt-tRNA and UTP decreased proportionally whereas that of mt-RNA increased. These results argue against different UTP pools during synthesis of cyt-rRNA and cyt-tRNA, but indicate compartmentation of UTP during rRNA synthesis in the nucleus and the mitochondria of tomato cells.Abbreviations CMP cytidine 5-monophosphate - cyt-rRNA cytoplasmic ribosomal RNA - cyt-tRNA cytoplasmic transfer RNA - mt-rRNA mitochondrial rRNA - NC nitrocellulose - PAGE polyacrylamide gel electrophoresis - TLC thin-layer chromatography - Tris 2-amino-2-(hydroxymethyl)-1,3-propanediol - UDP uridine 5-diphosphate - UMP uridine 5-monophosphate - UTP uridine 5-triphosphate     

Antisense RNAs everywhere?

In recent years, systematic searches of both prokaryote and eukaryote genomes have identified a staggering number of small RNAs, the biological functions of which remain unknown. Small RNA-based regulators are well known from bacterial plasmids. They act on target RNAs by sequence complementarity; that is, they are antisense RNAs. Recent findings suggest that many of the novel orphan RNAs encoded by bacterial and eukaryotic chromosomes might also belong to a ubiquitous, heterogeneous class of antisense regulators of gene expression.     

Regulatory small RNAs from the 3′ regions of bacterial mRNAs

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Critical roles of non-coding RNAs in lifecycle and biology of Marek’s disease herpesvirus

Science China Life Sciences - Over the past two decades, numerous non-coding RNAs (ncRNAs) have been identified in different biological systems including virology, especially in large DNA viruses...     

植物小RNAs的研究进展

小RNAs(长度小于40 nt)是nc-RNAs重要的一部分,现在植物中已发现了多种小RNAs,如小干扰RNAs(siRNAs)、微小RNAs(miRNAs)、反式作用的小干扰RNAs、天然反义转录小干扰RNAs、异染色质小干扰RNAs、长小片段小干扰RNAs、天然反义转录的微小RNAs及其一些未命名的小RNAs.成熟的小RNAs聚集相关的蛋白质因子,可以抑制转录,导致转录水平的基因沉默(TGS);或介导目标mRNA的剪切,抑制翻译,导致转录后水平基因沉默(PTGS).就这些植物小RNAs产生及其作用的研究进展作一概述     

抗流感病毒小RNAs研究进展

流行性感冒是一类由流感病毒引起的呼吸道传染病, 通过季节性流行或全球性爆发严重威胁着人类健康。目前防治流感的主要方法是疫苗和药物, 但存在神经毒性、肠胃副作用、易耐药等诸多限制因素。新的技术特别是小RNAs介导的RNA干扰(RNAi)技术, 因其具有高效、特异、快速等特点, 已成为抗病毒治疗的候选方法之一。随着近年来流感病毒的流行, 应用小RNAs抗流感病毒的报导越来越多, 其中靶向PA、NP和M2的PA-2087, NP-1496和M-950是目前报道的抑制流感病毒效果最好的siRNA。靶向不同流感病毒基因保守区域的siRNA具有更广泛的病毒毒株抑制效果, 靶向不同基因的siRNAs联合使用可取得更好的病毒抑制效果。文章就目前siRNAs和miRNAs在抗流感病毒方面的研究进展及RNAi治疗的前景和问题进行了综述。     

Modified 5′-termini in small nuclear RNAs of mouse myeloma cells

Nuclei of MPC 11 mouse myeloma cells contain several species of small RNAs related to those found in other mammalian cells. These include U1 RNA, about 190 nucleotides in length and U2 RNA, about 170 nucleotides long. The 5'-termini of 32P-labelled U1 and U2 RNAs have been investigated by a fingerprinting technique involving digestion with T2-ribonuclease. The RNAs were found to have modified 5'-terminal structures of the form m3G(5')ppp (5')AmpUmpAp for U1 RNA and m3G(5')ppp(5')AmpUmpCmpCp for U2 RNA, where m3G is N2, N27-trimethyl guanosine and Am and Um are 2'-O-methyl nucleosides. These 5'-terminal sequences are the same as those proposed for rat hepatoma U1 and U2 RNAs (Ro-Choi et al., Fed. Proc. 33, 1548, 1974) but with triphosphate rather than diphosphate links.     

抗流感病毒小RNAs研究进展

流行性感冒是一类由流感病毒引起的呼吸道传染病,通过季节性流行或全球性爆发严重威胁着人类健康。目前防治流感的主要方法是疫苗和药物,但存在神经毒性、肠胃副作用、易耐药等诸多限制因素。新的技术特别是小RNAs介导的RNA干扰(RNAi)技术,因其具有高效、特异、快速等特点,已成为抗病毒治疗的候选方法之一。随着近年来流感病毒的流行,应用小RNAs抗流感病毒的报导越来越多,其中靶向PA、NP和M2的PA-2087,NP-1496和M-950是目前报道的抑制流感病毒效果最好的siRNA。靶向不同流感病毒基因保守区域的siRNA具有更广泛的病毒毒株抑制效果,靶向不同基因的siRNAs联合使用可取得更好的病毒抑制效果。文章就目前siRNAs和miRNAs在抗流感病毒方面的研究进展及RNAi治疗的前景和问题进行了综述。     

A two-dimensional gel electrophoretic procedure for the separation of complex mixtures of 4–12S RNAs

A two-dimensional gel electrophoretic method suitable for the separation of complex mixtures of RNA species in the size range of 4 to 12 S is described. A 3.6–11% polyacrylamide gradient gel containing a gradient of 0–7 m urea was used in the first dimension, and a transverse 3.6–22.6% polyacrylamide gradient gel containing 5 m urea was used in the second dimension. The method was applied to the separation of total cytoplasmic RNAs from a cellular slime mold. In this method reproducible fingerprints were obtained by the use of visible-marker RNA.