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Nucleic Acids Research, 1991, Vol. 19, No. 23 6519-6525
© 1991

MOLECULAR BIOLOGY

Self-cleavage of a 71 nucleotide-long ribozyme derived from hepatitis delta virus genomic RNA

Gilbert Thill1, Marta Blumenfeld1, Franck Lescure1,2 and Marc Vasseur1,2,*

1GENSET, 1 Passage Etienne Delaunay 75011 Prais 2Laboratorie de Virologie Moleéculaire, Universiteé Paris VII 2 place Jussieu, 75251 Paris ceèdex 05, France

*To whom correspondence should be addressed at Laboratorie de Virologie Moleè culaire, Universiteè paris VII, 2 place Jussieu, 75251 Parisa Ceèdex 05, France

Received August 12, 1991. Accepted November 8, 1991.

Self-cleavage efficiency of ribozymes derived from hepatitis delta virus (HDV) has been shown to depend on the RNA structure, which in turn may be determined by the length of the considered sequences. Here we describe the construction and functional analysis of a 71 nucleotide-long RNA genomic fragment, Rz71, which carries an 18 nucleotide deletion in a very stable GC-rich stem-loop (stem IV), predicted to be present in several computer-derived secondary structure models. Rz71 is able to undergo self-cleavage under non-denaturing conditions (the t1/2of the reaction at 37°C is 3 min). The deletion, however, is not neutral, since under the same conditions the non-deleted ribozyme cleaves to 50% in less than 15 sec. Therefore, stem-loop IV seems to play a structural role, not being directly involved in the catalytic reaction, but contributing to the correct positioning of the catalytic core of the HDV ribozyme. Rz71 is the smallest selfcleaving sub-fragment of HDV genomic RNA reported so far.


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