Spatially restricted microRNA directs leaf polarity through ARGONAUTE1

Gene regulation by RNA interference requires the functions of the PAZ domain protein Argonaute. In plants, mutations in ARGONAUTE1 (AGO1) are associated with distinctive developmental defects that suggest a role for microRNA (miRNA) in organ polarity. Potential targets of miRNA regulation are the homeodomain/leucine zipper genes PHABULOSA (PHB) and PHAVOLUTA (PHV). These genes are expressed in a polar fashion in leaf primordia and are required for adaxial cell fate. Here we show that a 21-nucleotide miRNA that directs cleavage of PHB/PHV messenger RNA accumulates first in the embryonic meristem, and then in the abaxial domain of the developing leaf. miRNA distribution is disrupted by mutations in AGO1, indicating that AGO1 affects the regulation of miRNA. In addition, interactions between homeodomain/leucine zipper genes and an allelic series of ago1 indicate that miRNA acts as a signal to specify leaf polarity.     

SERRATE coordinates shoot meristem function and leaf axial patterning in Arabidopsis

Leaves of flowering plants are determinate organs produced by pluripotent structures termed shoot apical meristems. Once specified, leaves differentiate an adaxial (upper) side specialized for light capture, and an abaxial (lower) side specialized for gas exchange. A functional relationship between meristem activity and the differentiation of adaxial leaf fate has been recognized for over fifty years, but the molecular basis of this interaction is unclear. In Arabidopsis thaliana, activity of the class I KNOX (KNOTTED1-like homeobox) genes SHOOTMERISTEMLESS (STM) and BREVIPEDICELLUS (BP) is required for meristem function but excluded from leaves, whereas members of the HD-Zip III (class III homeodomain leucine zipper) protein family function to promote both meristem activity and adaxial leaf fate. Here we show that the zinc-finger protein SERRATE acts in a microRNA (miRNA) gene-silencing pathway to regulate expression of the HD-Zip III gene PHABULOSA (PHB) while also limiting the competence of shoot tissue to respond to KNOX expression. Thus, SERRATE acts to coordinately regulate meristem activity and leaf axial patterning.     

Role of PHABULOSA and PHAVOLUTA in determining radial patterning in shoots

The upper side of the angiosperm leaf is specialized for efficient capture of sunlight whereas the lower side is specialized for gas exchange. In Arabidopsis, the establishment of polarity in the leaf probably requires the generation and perception of positional information along the radial (adaxial versus abaxial or central versus peripheral) dimension of the plant. This is because the future upper (adaxial) side of the leaf develops from cells closer to the centre of the shoot, whereas the future under (abaxial) side develops from cells located more peripherally. Here we implicate the Arabidopsis PHABULOSA and PHAVOLUTA genes in the perception of radial positional information in the leaf primordium. Dominant phabulosa (phb) and phavoluta (phv) mutations cause a dramatic transformation of abaxial leaf fates into adaxial leaf fates. They do so by altering the predicted sterol/lipid-binding domains of ATHB14 and ATHB9, proteins of previously unknown function that also contain DNA-binding motifs. This change probably renders the protein constitutively active, implicating this domain as a central regulator of protein function and the PHB and PHV proteins as receptors for an adaxializing signal.     

水稻低温胁迫响应 ｍｉｃｒｏＲＮＡ 鉴定

水稻是世界三大粮食作物之一,然而低温胁迫会严重抑制水稻的生长发育。为了探究micoRNA在水稻低温胁迫中的作用,采用低温处理前,5℃低温处理24h和5℃低温处理48h的2~3叶期水稻整株,构建9个小RNA文库。通过高通量测序后,对9个小RNA文库的microRNA进行差异表达分析,一共筛选出21个与冷胁迫相关的microRNA,其中16个在冷胁迫下上调,5个在冷胁迫下下调。通过对这21个microRNA靶基因的CO富集结果表明,其靶基因广泛富集在包括信号转导,免疫系统和物质合成等细胞内过程中。这表明水稻可能通过多种micoRNA 介导,从各个方面来协同抵御低温胁迫。本研究为进一步阐明microRNA响应低温胁迫的分子机制提供了基础,且本研究所鉴定的microRNA为增强水稻对低温耐受性遗传改良提供了优异的miRNA资源。     

The functions of animal microRNAs

MicroRNAs (miRNAs) are small RNAs that regulate the expression of complementary messenger RNAs. Hundreds of miRNA genes have been found in diverse animals, and many of these are phylogenetically conserved. With miRNA roles identified in developmental timing, cell death, cell proliferation, haematopoiesis and patterning of the nervous system, evidence is mounting that animal miRNAs are more numerous, and their regulatory impact more pervasive, than was previously suspected.     

A viral microRNA functions as an orthologue of cellular miR-155

All metazoan eukaryotes express microRNAs (miRNAs), roughly 22-nucleotide regulatory RNAs that can repress the expression of messenger RNAs bearing complementary sequences. Several DNA viruses also express miRNAs in infected cells, suggesting a role in viral replication and pathogenesis. Although specific viral miRNAs have been shown to autoregulate viral mRNAs or downregulate cellular mRNAs, the function of most viral miRNAs remains unknown. Here we report that the miR-K12-11 miRNA encoded by Kaposi's-sarcoma-associated herpes virus (KSHV) shows significant homology to cellular miR-155, including the entire miRNA 'seed' region. Using a range of assays, we show that expression of physiological levels of miR-K12-11 or miR-155 results in the downregulation of an extensive set of common mRNA targets, including genes with known roles in cell growth regulation. Our findings indicate that viral miR-K12-11 functions as an orthologue of cellular miR-155 and probably evolved to exploit a pre-existing gene regulatory pathway in B cells. Moreover, the known aetiological role of miR-155 in B-cell transformation suggests that miR-K12-11 may contribute to the induction of KSHV-positive B-cell tumours in infected patients.     

Regulation of microRNA on plant development and viral infection

THE FIRST MIRNA WAS IDENTIFIED IN C. ELEGANS AS EARLY AS IN 1993; THE IMPORTANCE OF MIRNAS, HOWEVER, IS RECOGNIZED ONLY RECENTLY AFTER THE DISCOVERY OF MIRNAS EXISTING UNIVERSALLY IN EUKARYOTIC ORGANISMS. THE SECOND MIRNA WAS IDENTIFIED IN 2000[1]. SINCE …     

基于Gene Ontology的MicroRNA功能聚类

MicroRNA(miRNA)的许多生物过程是通过影响靶基因的转录后表达.miRNA与靶标之间的互补程度和性质决定其基因调控作用.结构相似性可以作为一个强有力的方法推断分子功能的相似性.然而,结构比对的方法来度量miRNA之间的相似性通常不太准确,而且时间开销大.对这些表达差异的miRNA的靶标基因进行聚类,可以很好地理解miRNA的功能.提出一个新的GO(gene Ontology)语义相似性的方法来区分miRNA功能组.该方法采用项信息和边的权重来度量GO项的权重.此外,2个GO图的共同项和非共同项还被用来度量这2个图之间的相似度.对于2个miRNA,它们之间的相似性可以用它们靶标基因标注的GO项的相似性来计算.实验结果表明此方法不仅可以将相似功能的miRNA聚在一起,而且可以预测未知miRNA的功能.     

Widespread changes in protein synthesis induced by microRNAs

Animal microRNAs (miRNAs) regulate gene expression by inhibiting translation and/or by inducing degradation of target messenger RNAs. It is unknown how much translational control is exerted by miRNAs on a genome-wide scale. We used a new proteomic approach to measure changes in synthesis of several thousand proteins in response to miRNA transfection or endogenous miRNA knockdown. In parallel, we quantified mRNA levels using microarrays. Here we show that a single miRNA can repress the production of hundreds of proteins, but that this repression is typically relatively mild. A number of known features of the miRNA-binding site such as the seed sequence also govern repression of human protein synthesis, and we report additional target sequence characteristics. We demonstrate that, in addition to downregulating mRNA levels, miRNAs also directly repress translation of hundreds of genes. Finally, our data suggest that a miRNA can, by direct or indirect effects, tune protein synthesis from thousands of genes.     

microRNA对肿瘤细胞增殖与分化的调控

microRNA(miRNA)是一种长度约为22核苷酸(nt)的非编码RNA,其主要通过碱基互补与靶mRNA的3'端非翻译区(3'UTR)结合,导致靶mRNA降解或抑制蛋白质的合成,在转录后水平调节基因的表达.miRNA突变、缺失或表达水平的异常会导致生理的异常与疾病的发生,与人类肿瘤疾病密切相关,它具有类似于癌基因或抑癌基因的作用,可参与肿瘤细胞的增殖、分化和细胞凋亡等调控过程.miRNA在肿瘤诊断和治疗方面具有广阔的应用前景.     

Silencing of microRNAs in vivo with 'antagomirs'

MicroRNAs (miRNAs) are an abundant class of non-coding RNAs that are believed to be important in many biological processes through regulation of gene expression. The precise molecular function of miRNAs in mammals is largely unknown and a better understanding will require loss-of-function studies in vivo. Here we show that a novel class of chemically engineered oligonucleotides, termed 'antagomirs', are efficient and specific silencers of endogenous miRNAs in mice. Intravenous administration of antagomirs against miR-16, miR-122, miR-192 and miR-194 resulted in a marked reduction of corresponding miRNA levels in liver, lung, kidney, heart, intestine, fat, skin, bone marrow, muscle, ovaries and adrenals. The silencing of endogenous miRNAs by this novel method is specific, efficient and long-lasting. The biological significance of silencing miRNAs with the use of antagomirs was studied for miR-122, an abundant liver-specific miRNA. Gene expression and bioinformatic analysis of messenger RNA from antagomir-treated animals revealed that the 3' untranslated regions of upregulated genes are strongly enriched in miR-122 recognition motifs, whereas downregulated genes are depleted in these motifs. Analysis of the functional annotation of downregulated genes specifically predicted that cholesterol biosynthesis genes would be affected by miR-122, and plasma cholesterol measurements showed reduced levels in antagomir-122-treated mice. Our findings show that antagomirs are powerful tools to silence specific miRNAs in vivo and may represent a therapeutic strategy for silencing miRNAs in disease.