The 5′-Terminal Nucleotide Sequences of the Double-Stranded RNA of Human Reovirus

AUTOR(ES)

Miura, Kin-Ichiro

RESUMO

The 5′-terminal nucleotide sequences of human reovirus double-stranded RNA were determined after labeling the RNA with [32P]phosphate by polynucleotide kinase. The 5′ terminal were labeled to only a limited extent prior to sequential oxidation, β-elimination, and phosphomonoesterase treatment, indicating that the terminal phosphates were in a modified, blocked configuration. Each genome segment, after removing the blocking group, contained the same two 5′-terminal sequences: GpApUp in one chain and G*pCp in the other. G*p is a derivative of guanylic acid, probably 2′-O-methyl-Gp, which renders the 5′-terminal sequence resistant to hydrolysis by alkali. The results indicate that the transcription of reovirus double-stranded RNA strats from the 3′ end complementary to the G*pCp-terminal, resulting in the synthesis of single-stranded mRNA carrying the same 5′ sequence as the G*pCp-chain. The presence of a modified nucleotide at the 5′ terminus of the strand complementary to the mRNA template is a feature common to another double-stranded RNA virus, cytoplasmic polyhedrosis virus.

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