在P1/HC-Pro拟南芥中异位表达DICER-LIKE1可挽救表型异常,但不能挽救微小RNA和沉默途径中的缺陷。

Ectopic DICER-LIKE1 expression in P1/HC-Pro Arabidopsis rescues phenotypic anomalies but not defects in microRNA and silencing pathways.

作者信息

Mlotshwa Sizolwenkosi, Schauer Stephen E, Smith Trenton H, Mallory Allison C, Herr J M, Roth Braden, Merchant Delwin S, Ray Animesh, Bowman Lewis H, Vance Vicki B

机构信息

Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208, USA.

出版信息

Plant Cell. 2005 Nov;17(11):2873-85. doi: 10.1105/tpc.105.036608. Epub 2005 Oct 7.

Abstract

Expression of the viral silencing suppressor P1/HC-Pro in plants causes severe developmental anomalies accompanied by defects in both short interfering RNA (siRNA) and microRNA (miRNA) pathways. P1/HC-Pro transgenic lines fail to accumulate the siRNAs that mediate RNA silencing and are impaired in both miRNA processing and function, accumulating abnormally high levels of miRNA/miRNA* processing intermediates as well as miRNA target messages. Both miRNA and RNA silencing pathways require participation of DICER-LIKE (DCL) ribonuclease III-like enzymes. Here, we investigate the effects of overexpressing DCL1, one of four Dicers in Arabidopsis thaliana, on P1/HC-Pro-induced defects in development and small RNA metabolism. Expression of a DCL1 cDNA transgene (35S:DCL1) produced a mild gain-of-function phenotype and largely rescued dcl1 mutant phenotypes. The 35S:DCL1 plants were competent for virus-induced RNA silencing but were impaired in transgene-induced RNA silencing and in the accumulation of some miRNAs. Ectopic DCL1 largely alleviated developmental anomalies in P1/HC-Pro plants but did not correct the P1/HC-Pro-associated defects in small RNA pathways. The ability of P1/HC-Pro plants to suppress RNA silencing and the levels of miRNAs, miRNA*s, and miRNA target messages in these plants were essentially unaffected by ectopic DCL1. These data suggest that P1/HC-Pro defects in development do not result from general impairments in small RNA pathways and raise the possibility that DCL1 participates in processes in addition to miRNA biogenesis.

摘要

病毒沉默抑制因子P1/HC-Pro在植物中的表达会导致严重的发育异常,并伴有小干扰RNA(siRNA)和微小RNA(miRNA)途径的缺陷。P1/HC-Pro转基因株系无法积累介导RNA沉默的siRNA,在miRNA加工和功能方面均受损,积累了异常高水平的miRNA/miRNA加工中间体以及miRNA靶标信息。miRNA和RNA沉默途径都需要类似Dicer的核糖核酸酶III(DCL)的参与。在此,我们研究了拟南芥四种Dicer之一的DCL1过表达对P1/HC-Pro诱导的发育缺陷和小RNA代谢的影响。DCL1 cDNA转基因(35S:DCL1)的表达产生了轻微的功能获得表型,并在很大程度上挽救了dcl1突变体表型。35S:DCL1植株具有病毒诱导的RNA沉默能力,但在转基因诱导的RNA沉默和一些miRNA的积累方面存在缺陷。异位表达的DCL1在很大程度上缓解了P1/HC-Pro植株的发育异常,但没有纠正P1/HC-Pro相关的小RNA途径缺陷。P1/HC-Pro植株抑制RNA沉默的能力以及这些植株中miRNA、miRNA和miRNA靶标信息的水平基本上不受异位DCL1的影响。这些数据表明,P1/HC-Pro的发育缺陷并非由小RNA途径的普遍损伤所致,并增加了DCL1除参与miRNA生物合成外还参与其他过程的可能性。

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