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  • Title: 3'-terminal nucleotide sequences important for the accumulation of cowpea mosaic virus M-RNA.
    Author: Rohll JB, Holness CL, Lomonossoff GP, Maule AJ.
    Journal: Virology; 1993 Apr; 193(2):672-9. PubMed ID: 8460480.
    Abstract:
    The location of nucleotide sequences important in determining the extent of cowpea mosaic virus M-RNA accumulation in cowpea protoplasts has been analyzed by deletion mutagenesis of full-length cDNA clones from which infectious transcripts can be produced in vitro. The results suggest that cis-acting sequences which direct replication of M-RNA by B-RNA-encoded products are located within the 5'-terminal 524 nucleotides and the 3'-terminal 151 nucleotides. RNA secondary structure predictions for the 3'-terminal 151 nucleotides of both genomic RNAs (Eggen et al. (1989) Virology 173, 456-464) indicate that the terminal nucleotides form a stable secondary structure composed of a Y-shaped stem-loop and a simple A-U-rich stem-loop. The latter structure has been implicated in B-RNA replication. We have examined the role of the Y-shaped structure in M-RNA accumulation by site-directed mutagenesis of putative base-pairing combinations in the two minor stems. The results suggest that efficient replication is dependent on the formation of both of these minor stem structures.
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