Red clover necrotic mosaic virus replication proteins accumulate at the endoplasmic reticulum.
Red clover necrotic mosaic virus (RCNMV) encodes N-terminally overlapping proteins of 27 and 88 kDa (p27 and p88) known to be required for replication. Green fluorescent protein (GFP) fusions were used to visualize the location of p27 and p88 within Nicotiana benthamiana cells. GFP:p27 fusions localized to the endoplasmic reticulum (ER), co-localized with ER-targeted yellow fluorescent protein and caused membrane restructuring and proliferation. Cellular fractionation of virus-inoculated N. benthamiana leaves confirmed the association of p27 with ER membranes. GFP:p88 fusions also localized to the ER and co-localized with GFP:p27. Both fusion proteins co-localize to the cortical and cytoplasmic ER and were associated with invaginations of the nuclear envelope. Independent accumulation in, and perturbation of, the ER suggests that p27 and p88 function together in the replication complex. This is the first report of a member of the Tombusviridae replicating in association with the ER.
Turner
Katherine A
KA
Department of Genetics, North Carolina State University, Raleigh, NC 27695-7614, USA.
Sit
Tim L
TL
Callaway
Anton S
AS
Allen
Nina S
NS
Lommel
Steven A
SA
D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000821:virology; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D009124:Muscle Proteins; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000821:virology; D019183:Tombusviridae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D029921:Trifolium / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D008164:Luminescent Proteins; D008840:Microfilament Proteins; D009124:Muscle Proteins; D011993:Recombinant Fusion Proteins; C058992:Tagln protein, mouse; D014764:Viral Proteins; D049452:Green Fluorescent Proteins