Identification of a sequence element in the alphavirus core protein which mediates interaction of cores with ribosomes and the disassembly of cores.
Early in infection core protein is transferred from alphavirus cores to ribosomes (Wengler and Wengler, 1984, Virology 134, 435-442) and it has been suggested that ribosome binding is a property of alphavirus core protein which is involved in core disassembly. Here we describe in vitro analyses of this transfer. Sindbis virus cores, incubated with ribosomes either in a reticulocyte lysate or in buffer, are disassembled with a concomitant transfer of core protein to the large ribosomal subunit. Preincubation of ribosomes with core protein blocks disassembly. Limited proteolysis of Sindbis virus core releases the carboxy-terminal core protein domain as a soluble fragment (Strong and Harrison, 1990, J. Virol. 64, 3992-3994). Trypsin- or proteinase Lys-C-released fragments contain the amino-terminal residue met (106) or gln (94), respectively. The fragment generated by proteinase Lys-C binds to ribosomes and interferes with core disassembly whereas the slightly shorter tryptic fragment has none of these activities. These and further analyses indicate that a conserved sequence element which surrounds amino acid met (106) of SIN CP, the so-called RBSc element, leads to binding of core protein to ribosomes and thereby to core disassembly. Implications of the experiments for regulation of assembly of alphavirus cores and for the core protein-induced resistance to viral multiplication observed in plant virus systems are discussed.
Wengler
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G
Institut für Virologie, Justus-Liebig-Universität Giessen, Germany.
Würkner
D
D
Wengler
G
G
D000524:Alphavirus / Q000235:genetics; D000595:Amino Acid Sequence; D001665:Binding Sites; D010450:Endopeptidases / Q000494:pharmacology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010449:Peptide Mapping; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D012845:Sindbis Virus / Q000737:chemistry / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014758:Viral Core Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D010446:Peptide Fragments; D014758:Viral Core Proteins; D010450:Endopeptidases