RNA-dependent RNA polymerase activity associated with endogenous double-stranded RNA in rice

Plant Cell Physiol. 2001 Feb;42(2):197-203. doi: 10.1093/pcp/pce025.

Abstract

RNA-dependent RNA polymerase (RdRp) activity was detected in the crude microsomal fraction of rice cultured cells that contain a 14 kbp double-stranded RNA (dsRNA). RdRp activity is maximal in the presence of all four nucleotide triphosphates and Mg2+ ion and is resistant to inhibitors of DNA-dependent RNA polymerases (actinomycin D and alpha-amanitin). RdRp activity increases approximately 2.5-fold in the presence of 0.5% deoxycholate. Treatment of purified microsomal fraction with proteinase K plus deoxycholate suggests that the RdRp enzyme complex with its own 14 kb RNA template is located in vesicles. The RdRp enzyme complex was solubilized with Nonidet P-40 and purified by glycerol gradient centrifugation, then exogenous RNA templates were added. Results indicate that exogenous dsRNA reduces RNA synthesis from the endogenous 14 kb RNA template.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Centrifugation, Density Gradient
  • Detergents / pharmacology
  • Microsomes / drug effects
  • Oryza / enzymology*
  • Oryza / genetics
  • RNA, Double-Stranded / metabolism*
  • RNA, Plant / metabolism*
  • RNA-Dependent RNA Polymerase / metabolism*
  • Sucrose

Substances

  • Detergents
  • RNA, Double-Stranded
  • RNA, Plant
  • Sucrose
  • RNA-Dependent RNA Polymerase