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首页> 外文期刊>PLoS Genetics >Evidence for Antisense Transcription Associated with MicroRNA Target mRNAs in Arabidopsis
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Evidence for Antisense Transcription Associated with MicroRNA Target mRNAs in Arabidopsis

机译：与拟南芥中MicroRNA靶mRNA相关的反义转录的证据。

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Antisense transcription is a pervasive phenomenon, but its source and functional significance is largely unknown. We took an expression-based approach to explore microRNA (miRNA)-related antisense transcription by computational analyses of published whole-genome tiling microarray transcriptome and deep sequencing small RNA (smRNA) data. Statistical support for greater abundance of antisense transcription signatures and smRNAs was observed for miRNA targets than for paralogous genes with no miRNA cleavage site. Antisense smRNAs were also found associated with MIRNA genes. This suggests that miRNA-associated “transitivity” (production of small interfering RNAs through antisense transcription) is more common than previously reported. High-resolution (3 nt) custom tiling microarray transcriptome analysis was performed with probes 400 bp 5′ upstream and 3′ downstream of the miRNA cleavage sites (direction relative to the mRNA) for 22 select miRNA target genes. We hybridized RNAs labeled from the smRNA pathway mutants, including hen1-1, dcl1-7, hyl1-2, rdr6-15, and sgs3-14. Results showed that antisense transcripts associated with miRNA targets were mainly elevated in hen1-1 and sgs3-14 to a lesser extent, and somewhat reduced in dcl11-7, hyl11-2, or rdr6-15 mutants. This was corroborated by semi-quantitative reverse transcription PCR; however, a direct correlation of antisense transcript abundance in MIR164 gene knockouts was not observed. Our overall analysis reveals a more widespread role for miRNA-associated transitivity with implications for functions of antisense transcription in gene regulation. HEN1 and SGS3 may be links for miRNA target entry into different RNA processing pathways.
机译：反义转录是一种普遍现象，但是其来源和功能意义在很大程度上是未知的。通过对已发表的全基因组平铺微阵列转录组和深度测序小RNA（smRNA）数据进行计算分析，我们采用了基于表达的方法来探索与microRNA（miRNA）相关的反义转录。与没有miRNA切割位点的旁系同源基因相比，miRNA目标的反义转录签名和smRNA的丰度得到了更多的统计支持。还发现反义smRNA与MIRNA基因相关。这表明与miRNA相关的“传递性”（通过反义转录产生小的干扰RNA）比以前报道的更为普遍。使用miRNA裂解位点上游5'上游和3'下游400 bp的探针对22个选定的miRNA靶基因进行高分辨率（3 nt）定制切片微阵列转录组分析。我们杂交了从smRNA途径突变体（包括hen1-1，dcl1-7，hyl1-2，rdr6-15和sgs3-14）标记的RNA。结果显示，与miRNA靶标相关的反义转录本主要在hen1-1和sgs3-14中升高程度较小，而在dcl11-7，hyl11-2或rdr6-15突变体中则有所降低。半定量逆转录PCR证实了这一点。然而，未观察到MIR164基因敲除中反义转录物丰度的直接相关性。我们的整体分析显示，与miRNA相关的传递性具有更广泛的作用，对基因调控中反义转录的功能有影响。 HEN1和SGS3可能是miRNA靶标进入不同RNA加工途径的链接。

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