title: Analysis of the role of the coat protein N-terminal segment in Potato virus X virion stability and functional activity.
abstract: Previously, we have reported that intact Potato virus X (PVX) virions cannot be translated in cell-free systems, but acquire this capacity by the binding of PVX-specific triple gene block protein 1 (TGBp1) or after phosphorylation of the exposed N-terminal segment of intravirus coat protein (CP) by protein kinases. With the help of in vitro mutagenesis, a nonphosphorylatable PVX mutant (denoted ST PVX) was prepared in which all 12 S and T residues in the 20-residue-long N-terminal CP segment were substituted by A or G. Contrary to expectations, ST PVX was infectious, produced normal progeny and was translated in vitro in the absence of any additional factors. We suggest that the N-terminal PVX CP segment somehow participates in virion assembly in vivo and that CP subunits in ST virions may differ in structure from those in the wild-type (UK3 strain). In the present work, to test this suggestion, we performed a comparative tritium planigraphy study of CP structure in UK3 and ST virions. It was found that the profile of tritium incorporation into ST mutant virions in some CP segments differed from that of normal UK3 virions and from UK3 complexed with the PVX movement protein TGBp1. It is proposed that amino acid substitutions in ST CP and the TGBp1-driven remodelling of UK3 virions induce structural alterations in intravirus CPs. These alterations affect the predicted RNA recognition motif of PVX CP, but in different ways: for ST PVX, labelling is increased in α-helices 6 and 7, whereas, in remodelled UK3, labelling is increased in the β-sheet strands β3, β4 and β5.
authors: AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119992 Moscow, Russia.
authors: Elena
authors: E
authors: Lukashina
authors: Alexander
authors: A
authors: Ksenofontov
authors: Natalia
authors: N
authors: Fedorova
authors: Gennady
authors: G
authors: Badun
authors: Anna
authors: A
authors: Mukhamedzhanova
authors: Olga
authors: O
authors: Karpova
authors: Nina
authors: N
authors: Rodionova
authors: Lyudmila
authors: L
authors: Baratova
authors: Evgeny
authors: E
authors: Dobrov
mesh_terms: D000595:Amino Acid Sequence; D036022:Capsid Proteins / Q000737:chemistry* / Q000378:metabolism*; D007553:Isotope Labeling; D008969:Molecular Sequence Data; D017863:Potexvirus / Q000378:metabolism*; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D014316:Tritium / Q000378:metabolism; D014771:Virion / Q000378:metabolism*
chemical_list: D036022:Capsid Proteins; C058750:coat protein, Potato virus X; D014316:Tritium