Phosphorylation down-regulates the RNA binding function of the coat protein of potato virus A.
Plant viruses encode movement proteins (MPs) to facilitate transport of their genomes from infected into neighboring healthy cells through plasmodesmata. Growing evidence suggests that specific phosphorylation events can regulate MP functions. The coat protein (CP) of potato virus A (PVA; genus Potyvirus) is a multifunctional protein involved both in virion assembly and virus movement. Labeling of PVA-infected tobacco leaves with [(33)P]orthophosphate demonstrated that PVA CP is phosphorylated in vivo. Competition assays established that PVA CP and the well characterized 30-kDa MP of tobacco mosaic virus (genus Tobamovirus) are phosphorylated in vitro by the same Ser/Thr kinase activity from tobacco leaves. This activity exhibits a strong preference for Mn(2+) over Mg(2+), can be inhibited by micromolar concentrations of Zn(2+) and Cd(2+), and is not Ca(2+)-dependent. Tryptic phosphopeptide mapping revealed that PVA CP was phosphorylated by this protein kinase activity on multiple sites. In contrast, PVA CP was not phosphorylated when packaged into virions, suggesting that the phosphorylation sites are located within the RNA binding domain and not exposed on the surface of the virion. Furthermore, two independent experimental approaches demonstrated that the RNA binding function of PVA CP is strongly inhibited by phosphorylation. From these findings, we suggest that protein phosphorylation represents a possible mechanism regulating formation and/or stability of viral ribonucleoproteins in planta.
Ivanov
K I
KI
Institute of Biotechnology, Viikki Biocenter, P.O. Box 56, FIN-00014 University of Helsinki, Finland.
Puustinen
P
P
Merits
A
A
Saarma
M
M
Mäkinen
K
K
D001667:Binding, Competitive; D002104:Cadmium / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015151:Immunoblotting; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008280:Magnetics; D008345:Manganese / Q000378:metabolism; D010766:Phosphorylation; D010942:Plant Viruses / Q000737:chemistry*; D010947:Plants, Toxic; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D013912:Threonine / Q000378:metabolism; D014026:Nicotiana / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology; D015032:Zinc / Q000378:metabolism
D004791:Enzyme Inhibitors; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D002104:Cadmium; D013912:Threonine; D008345:Manganese; D012694:Serine; D012313:RNA; D014357:Trypsin; D019311:Staurosporine; D008274:Magnesium; D015032:Zinc; D002118:Calcium