The conserved, hydrophilic and arginine-rich N-terminal domain of cucumovirus coat proteins contributes to their anomalous electrophoretic mobilities in sodium dodecylsulfate-polyacrylamide gels.
Although the Mr values of the coat proteins (CPs) of several cucumoviruses have been calculated from their deduced amino acid sequences to be approximately 24,000, the experimentally determined M(r) values using the Laemmli SDS-PAGE system were 30,000-31,000. Examination of the amino acid composition revealed that these CPs are neither highly acidic nor highly basic. Post-translational glycosylation or phosphorylation were also ruled out as contributing factors to the observed anomalous electrophoretic mobility because the products of in vitro translation of cucumovirus RNA 4 and in vivo bacterial expression of the cloned CP gene co-migrated with authentic cucumovirus CPs. Comparison of the hydropathy profiles of the CPs revealed the presence in each of a strikingly similar, highly hydrophilic N-terminal domain of 30-32 amino acid residues that contains a cluster of basic amino acids, mainly arginine. Selective chemical cleavage at tryptophan residues in the CPs of cucumoviruses, known to contain single tryptophan residues, yielded two peptides; an N-terminal peptide that contained the conserved hydrophilic domain and a C-terminal peptide. SDS-PAGE analysis showed that the N-terminal, but not the C-terminal, peptide exhibited the anomalous electrophoretic mobility.
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Department of Plant Pathology, University of Kentucky, Lexington 40546, USA.
Ghabrial
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D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry* / Q000235:genetics; D036022:Capsid Proteins; D017124:Conserved Sequence; D017799:Cucumovirus / Q000737:chemistry* / Q000235:genetics; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017386:Sequence Homology, Amino Acid; D014867:Water
D036022:Capsid Proteins; D004279:DNA, Viral; D014867:Water; C070340:coat protein, peanut stunt virus; D001120:Arginine