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Nucleic Acids Research, 1985, Vol. 13, No. 21 7881-7897
© 1985

Articles

DNA and RNA polymerase activities of nuclei and hypotonic extracts of nuclei isolated from tomato golden mosaic virus infected tobacco leaves

R.H.A. Coutts+ and K.W. Buck

Department of Pure and Applied Biology, Imperial College of Science and Technology London SW7 2BB, UK

+Author to whom reprint requests should be sent

Received August 6, 1985. Revised October 7, 1985. Accepted October 7, 1985.

Nuclei and hypotonically leached extracts of nuclei prepared from tomato golden mosaic virus (TGMV)-infected Nicotiana benthamiana leaves have been used in in vitro DNA and RNA polymerisation reactions. The synthesis of virus-specific DNA was resistant to aphidicolin, sensitive to N-ethylmaleimide and dideoxy TTP, and stimulated by KCl and ATP. Variably virion (+) and complementary (–) strand DNA of both the A and B genomic components were synthesised. Virus-specific RNA was synthesised in reactions which were initiated prior to nuclei isolation and leaching. From inhibitor studies and salt requirements RNA synthesis appeared to be catalysed by a DNA-dependent RNA polynerase type II enzyme. Both components of the TGMV genome were transcribed in a bidirectional fashion with a prevalence in some experiments of transcripts derived from DNA component A.


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