Journal Article

RNA-Dependent RNA Polymerase Activity Associated with Endogenous Double-Stranded RNA in Rice

Hideki Horiuchi, Tsuyoshi Udagawa, Ryuichi Koga, Hiromitsu Moriyama and Toshiyuki Fukuhara

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 2, pages 197-203
Published in print February 2001 | ISSN: 0032-0781
Published online February 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce025
RNA-Dependent RNA Polymerase Activity Associated with Endogenous Double-Stranded RNA in Rice

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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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 α-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.

Keywords: Key words: Double-stranded RNA — Endornavirus — Rice (Oryza sativa L.) — RNA-dependent RNA polymerase — Vesicle.; Abbreviations: dsRNA, double-stranded RNA; NP-40, Nonidet P-40; nt, nucleotides; OSV, Oryza sativa endornavirus; RdRp, RNA-dependent RNA polymerase; TMV, tobacco mosaic virus.

Journal Article.  4725 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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