A novel function for a ubiquitous plant enzyme pectin methylesterase: the enhancer of RNA silencing.
Co-agroinjection of Nicotiana benthamiana leaves with the pectin methylesterase (proPME) gene and the TMV:GFP vector resulted in a stimulation of virus-induced RNA silencing (inhibition of GFP production, virus RNA degradation, stimulation of siRNAs production). Conversely, co-expression of TMV:GFP with either antisense PME construct or with enzymatically inactive proPME restored synthesis of viral RNA. Furthermore, expression of proPME enhanced the GFP transgene-induced gene silencing accompanied by relocation of the DCL1 protein from nucleus to the cytoplasm and activation of siRNAs and miRNAs production. It was hypothesized that DCL1 relocated to the cytoplasm may use as substrates both miRNA precursor and viral RNA. The capacity for enhancing the RNA silencing is a novel function for the polyfunctional PME.
Dorokhov
Yuri L
YL
Department of Virology and A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Leninsky Gory 1, Laboratory Building A, Moscow 119992, Russia.
Frolova
Olga Y
OY
Skurat
Eugene V
EV
Ivanov
Peter A
PA
Gasanova
Tatjana V
TV
Sheveleva
Anna A
AA
Ravin
Nikolay V
NV
Mäkinen
Kristiina M
KM
Klimyuk
Victor I
VI
Skryabin
Konstantin G
KG
Gleba
Yuri Y
YY
Atabekov
Joseph G
JG
D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004792:Enzyme Precursors; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000096:biosynthesis; D019441:Plant Epidermis / Q000166:cytology; D018515:Plant Leaves / Q000166:cytology / Q000382:microbiology; D010940:Plant Proteins / Q000378:metabolism; D021381:Protein Transport; D034622:RNA Interference; D020871:RNA Stability / Q000235:genetics; D034741:RNA, Small Interfering / Q000096:biosynthesis; D012367:RNA, Viral / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics; D014026:Nicotiana / Q000201:enzymology*; D014027:Tobacco Mosaic Virus / Q000502:physiology; D019076:Transgenes
D004792:Enzyme Precursors; D035683:MicroRNAs; D010940:Plant Proteins; D034741:RNA, Small Interfering; D012367:RNA, Viral; D049452:Green Fluorescent Proteins; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase