RNA-primed complementary-sense DNA synthesis of the geminivirus African cassava mosaic virus.
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.
Saunders
K
K
Department of Virus Research, John Innes Institute, John Innes Centre for Plant Science Research, Norwich, UK.
Lucy
A
A
Stanley
J
J
D004279:DNA, Viral / Q000096:biosynthesis*; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D005559:Formamides; D009029:Mosaic Viruses / Q000235:genetics*; D010947:Plants, Toxic; D012367:RNA, Viral / Q000378:metabolism*; D014026:Nicotiana
D004279:DNA, Viral; D005559:Formamides; D012367:RNA, Viral