Poliovirus RNA synthesis utilizes an RNP complex formed around the 5'-end of viral RNA.
The structure of a ribonucleoprotein complex formed at the 5'-end of poliovirus RNA was investigated. This complex involves the first 90 nucleotides of poliovirus genome which fold into a cloverleaf-like structure and interact with both uncleaved 3CD, the viral protease-polymerase precursor, and a 36 kDa ribosome-associated cellular protein. The cellular protein is required for complex formation and interacts with unpaired bases in one stem-loop of the cloverleaf RNA. Amino acids within the 3C protease which are important for RNA binding were identified by site-directed mutagenesis and the crystal structure of a related protease was used to model the RNA binding domain within the viral 3CD protein. The physiologic importance of the ribonucleic-protein complex is suggested by the finding that mutations that disrupt complex formation abolish RNA replication but do not affect RNA translation or stability. Based on these structural and functional findings we propose a model for the initiation of poliovirus RNA synthesis where an initiation complex consisting of 3CD, a cellular protein, and the 5'-end of the positive strand RNA catalyzes in trans the initiation of synthesis of new positive stranded RNA.
Andino
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Rockefeller University, New York, NY 10021.
Rieckhof
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Achacoso
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PL
Baltimore
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004926:Escherichia coli; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D017955:Poliovirus / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D012367:RNA, Viral / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000378:metabolism*
D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins