Novel miR390-dependent transacting siRNA precursors in plants revealed by a PCR-based experimental approach and database analysis

J Biomed Biotechnol. 2009:2009:952304. doi: 10.1155/2009/952304. Epub 2009 Oct 13.

Abstract

TAS loci in plant genomes encode transacting small interfering RNAs (ta-siRNAs) that regulate expression of a number of genes. The function of TAS3 precursor in Arabidopsis thaliana is controlled by two miR390 target sites flanking two ta-siARF sequences targeting mRNAs of ARF transcription factors. Cleavage of the 3'-miR390-site initiates ta-siRNAs biogenesis. Here we describe the new method for identification of plant ta-siRNA precursors based on PCR with oligodeoxyribonucleotide primers mimicking miR390. The method was found to be efficient for dicotiledonous plants, cycads, and mosses. Based on sequences of amplified loci and a database analysis, a novel type of miR390-dependent TAS sequences was identified in dicots. These TAS loci are characterized by a smaller distance between miR390 sites compared to TAS3, a single copy of ta-siARF, and a sequence conservation pattern pointing to the possibility that processing of novel TAS-like locus is initiated by cleavage of the 5'-terminal miR390 target site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Base Sequence
  • DNA Primers / genetics
  • DNA, Plant / genetics
  • Databases, Nucleic Acid
  • MicroRNAs / genetics*
  • Molecular Sequence Data
  • Nicotiana / genetics
  • Plants / genetics*
  • Polymerase Chain Reaction
  • RNA Precursors / genetics*
  • RNA, Plant / genetics*
  • RNA, Small Interfering / genetics*
  • Sequence Homology, Nucleic Acid
  • Species Specificity
  • Transcriptional Activation

Substances

  • DNA Primers
  • DNA, Plant
  • MicroRNAs
  • RNA Precursors
  • RNA, Plant
  • RNA, Small Interfering