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Nucleic Acids Research, 1991, Vol. 19, No. 8 1893-1898
© 1991

MOLECULAR BIOLOGY

Alternative hammerhead structures in the self-cleavage of avocado sunblotch viroid RNAs

Christopher Davies§, Candice C. Sheldon§,+ and Robert H. Symons*,+

Department of Biochemistry, University of Adelaide SA 5001, Australia

* To whom correspondence should be addressed

Received November 12, 1990. Revised February 8, 1991. Accepted February 8, 1991.

The plus and minus RNAs of the 247 nt avocado sunblotch virold (ASBV) undergo site specific RNA self-cleavage reactions in vitro. As with several other self cleaving RNAs, we proposed hammerhead secondary structures for the sequence around the site of self-cleavage of both RNAs. We have shown previously that, during transcription of a dimeric plus ASBV RNA, a double-hammerhead structure formed and was necessary for self-cleavage. Here, we show that the purified full-length dimeric plus RNA, when incubated under our standard self-cleavage conditions, also self- cleaved by a double-hammerhead structure. In contrast, a dimeric minus ASBV RNA self-cleaved by a double-hammerhead structure during transcription, but by a single-hammerhead structure after purification. This illustrates the importance of the pathway of folding of the RNA in determining which active self-cleaving structure is formed.

+Present addresses: Department of Plant Science, Waite Instinite, Glen Osmond, SA 5064, Australia

§The Sainsbury Laboratory, John Innes Institute, Colney Lane, Norwich NR4 7UH, UK


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