Translational activation of encapsidated potato virus X RNA by coat protein phosphorylation.
Previously we showed that encapsidated potato virus X (PVX) RNA is nontranslatable in vitro, but can be converted into a translatable form after binding to PVX particles of PVX-coded movement protein, the product of the first gene of triple gene block (TGBp1). Here we report that a similar effect occurs via in situ phosphorylation of the PVX coat protein (CP) by Ser/Thr protein kinase (PK) C, the mixture of casein kinases I and II or by cytoplasmic PK(s) from Nicotiana glutinosa leaves. Immunochemical analyses indicated that phosphorylation induced conformational changes in PVX CP. The N-terminal region of the PVX CP, rich in Ser and Thr residues, is exposed at the virion surface and can be removed by treatment with trypsin. We showed that (i) trypsin treatment removed the bulk of (32)P-radioactivity from in situ phosphorylated PVX CP, (ii) PVX containing N-terminally truncated CP (PVX-Ptd) failed to be translationally activated by phosphorylation, and (iii) the specific infectivity of PVX-Ptd was reduced. However, the PVX-Ptd RNA remained intact and PVX-Ptd could be translationally activated by the PVX MP TGBp1. We hypothesize that phosphorylation of the parental PVX by cytoplasmic PK(s) in vivo renders PVX RNA translatable in primary inoculated cells, whereas translational activation of the progeny virions destined for plasmodesmata trafficking is triggered by TGBp1.
Atabekov
J G
JG
Department of Virology and A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia.
Rodionova
N P
NP
Karpova
O V
OV
Kozlovsky
S V
SV
Novikov
V K
VK
Arkhipenko
M V
MV
D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017863:Potexvirus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011198:Solanum tuberosum / Q000821:virology; D015533:Transcriptional Activation
D012367:RNA, Viral