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Nucleic Acids Research, 1992, Vol. 20, No. 23 6311-6315
© 1992

MOLECULAR BIOLOGY

RNA-primed complementary-sense DNA synthesis of the geminivirus African cassava mosaic virus

Keith Saunders, Andrew Lucy and John Stanley

Department of Virus Research, John Innes Institute, John Innes Centre for Plant Science Research Colney Lane, Norwich NR4 7UH, UK

Received August 25, 1992. Revised November 10, 1992. Accepted November 10, 1992.

The plant DNA virus African cassava mosaic virus (ACMV) is believed to replicate by a rolling circle mechanism. To investigate complementary-sense DNA (lagging strand) synthesis, we have analysed the heterogenous form of complementary-sense DNA (H3 DNA) from infected Nicotiana benthamiana by two-dimensional agarose gel electrophoresis and blot hybridisation. The presence of an RNA moeity is demonstrated by comparison of results for nucleic acids resolved on neutral/alkaline and neutral/formamide gels, suggesting that complementary-sense DNA synthesis on the virus-sense single-stranded DNA template is preceded by the synthesis of an RNA primer. Hybridisation with probes to specific parts of ACMV DNA A genome indicates that synthesis of the putative RNA primer initiates between nucleotides 2581–221, a region that includes intergenic sequences that have been implicated in geminivirus DNA replication and the control of gene expression.


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