Virus-induced gene silencing of the RPC5-like subunit of RNA polymerase III caused pleiotropic effects in Nicotiana benthamiana.
In eukaryotic cells, RNA polymerase III is highly conserved and transcribes housekeeping genes such as ribosomal 5S rRNA, tRNA and other small RNAs. The RPC5-like subunit is one of the 17 subunits forming RNAPIII and its exact functional roles in the transcription are poorly understood. In this work, we report that virus-induced gene silencing of transcripts encoding a putative RPC5-like subunit of the RNA Polymerase III in a model species Nicotiana benthamiana had pleiotropic effects, including but not limited to severe dwarfing appearance, chlorosis, nearly complete reduction of internodes and abnormal leaf shape. Using transcriptomic analysis, we identified genes and pathways affected by RPC5 silencing and thus presumably related to the cellular roles of the subunit as well as to the downstream cascade of reactions in response to partial loss of RNA Polymerase III function. Our results suggest that silencing of the RPC5L in N. benthamiana disrupted not only functions commonly associated with the core RNA Polymerase III transcripts, but also more diverse cellular processes, including responses to stress. We believe this is the first demonstration that activity of the RPC5 subunit is critical for proper functionality of RNA Polymerase III and normal plant development.
USDA/ARS, Beltsville Agricultural Research Center, Molecular Plant Pathology Laboratory, Beltsville MD 20705, USA.
Lev G
LG
Nemchinov
Institute of Basic Biological Problems, Russian Academy of Sciences, 2 Institute Street, Pushchino, Moscow Region, 142292, Russia.
Alexander M
AM
Boutanaev
USDA/ARS, Beltsville Agricultural Research Center, Molecular Plant Pathology Laboratory, Beltsville MD 20705, USA.
Olga A
OA
Postnikova
D001483:Base Sequence; D018506:Gene Expression Regulation, Plant; D063990:Gene Ontology; D020868:Gene Silencing; D017343:Genes, Plant; D058685:Genetic Pleiotropy; D010641:Phenotype; D010788:Photosynthesis; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D010942:Plant Viruses / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012320:RNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D017423:Sequence Analysis, RNA; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics / Q000821:virology*; D059467:Transcriptome / Q000235:genetics
D010940:Plant Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; D012320:RNA Polymerase III