A biochemical framework for RNA silencing in plants
Tang, Guiliang ; Reinhart, Brenda J. ; Bartel, David P. ; Zamore, Phillip D.
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Keywords
Arabidopsis Proteins
Endoribonucleases
*Gene Expression Regulation, Plant
Genes, Dominant
Homeodomain Proteins
MicroRNAs
Plant Extracts
Plant Proteins
*RNA Interference
RNA Replicase
RNA, Double-Stranded
RNA, Plant
RNA, Small Interfering
Ribonuclease III
Triticum
Life Sciences
Medicine and Health Sciences
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Abstract
RNA silencing phenomena were first discovered in plants, yet only the RNA interference pathway in animals has been subject to biochemical analysis. Here, we extend biochemical analysis to plant RNA silencing. We find that standard wheat germ extract contains Dicer-like enzymes that convert double-stranded RNA (dsRNA) into two classes of small interfering RNAs, as well as an RNA-dependent RNA polymerase activity that can convert exogenous single-stranded RNA into dsRNA. In this plant embryo extract, an endogenous microRNA (miRNA) that lacks perfect complementarity to its RNA targets nonetheless acts as a small interfering RNA. The miRNA guides an endonuclease to cleave efficiently wild-type Arabidopsis PHAVOLUTA mRNA, but not a dominant mutant previously shown to perturb leaf development. This finding supports the view that plant miRNAs direct RNAi and that miRNA-specified mRNA destruction is important for proper plant development. Thus, endonuclease complexes guided by small RNAs are a common feature of RNA silencing in both animals and plants.
Source
Genes Dev. 2003 Jan 1;17(1):49-63. Link to article on publisher's site