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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabidopsis / genetics*
  • Arabidopsis / growth & development
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics*
  • Cell Cycle Proteins / genetics*
  • Cysteine Endopeptidases / genetics*
  • DNA, Complementary / genetics
  • DNA, Plant / genetics
  • Gene Expression Regulation, Plant / genetics
  • Gene Silencing / physiology*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Mutation / genetics
  • Phenotype
  • Plant Viruses / genetics*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / growth & development
  • Plants, Genetically Modified / metabolism
  • RNA, Plant / genetics
  • RNA, Plant / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Ribonuclease III / genetics*
  • Signal Transduction / genetics
  • Transgenes / genetics
  • Viral Proteins / genetics*

Substances

  • Arabidopsis Proteins
  • Cell Cycle Proteins
  • DNA, Complementary
  • DNA, Plant
  • MicroRNAs
  • RNA, Plant
  • RNA, Small Interfering
  • Viral Proteins
  • DCL1 protein, Arabidopsis
  • Ribonuclease III
  • Cysteine Endopeptidases
  • HC-Pro protein, potyvirus