Abstract
Sequence analysis of an intracellular defective-interfering (DI) RNA, DIssE, of mouse hepatitis virus (MHV) revealed that it is composed of three noncontiguous genomic regions, representing the first 864 nucleotides of the 5′-end, an internal 748 nucleotides of the polymerase gene, and 601 nucleotides from the 3′-end of the parental MHV genome. DIssE had three base substitutions within the leader sequence and also a deletion of nine nucleotides located at the junction of the leader and the remaining genomic sequence. A system was developed for generating DI RNAs to study the mechanism of MHV RNA replication. A cDNA copy of DIssE RNA was placed downstream of T7 RNA polymerase promoter to generate DI RNAs capable of extremely efficient replication in the presence of a helper virus. We demonstrated that, in the DI RNA-transfected cells, the leader sequence of these DI RNAs was switched to that of the helper virus during one round of replication. This high-frequency leader sequence exchange was not observed if a nine-nucleotide stretch at the junction between the leader and the remaining DI sequence was deleted. This observation suggests that a free leader RNA is utilized for the replication of MHV RNA.
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© 1990 Plenum Press, New York
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Makino, S., Lai, M.M.C. (1990). Studies of Coronavirus DI RNA Replication Using In Vitro Constructed DI cDNA Clones. In: Cavanagh, D., Brown, T.D.K. (eds) Coronaviruses and their Diseases. Advances in Experimental Medicine and Biology, vol 276. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5823-7_46
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DOI: https://doi.org/10.1007/978-1-4684-5823-7_46
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