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Nucleic Acids Research, 1993, Vol. 21, No. 8 1713-1717
© 1993

MOLECULAR BIOLOGY

Translational control by a long range RNA–RNA interaction; a basepair substitution analysis

J. van Himbergen, B. van Geffen and J. van Duin*

Department of Biochemistry, Leiden University, Gorlaeus Laboratories PO Box 9502, 2300 RA Leiden, The Netherlands

* To whom correspondence should be addressed

Received February 22, 1993. Accepted March 23, 1993.

One of the two mechanisms that regulate expression of the repllcase clstron In the single stranded RNA collphages is translatlonal coupling. This mechanism prevents rlbosomes from binding at the start of the repllcase clstron unless the upstream coat clstron Is being translated. Genetic analysis had identified a maximal region of 132 nucleotides In the coat gene over which rlbosomes should pass to activate the replicase start. Subsequent deletion studies in our laboratory had further narrowed down the regulatory region to 12 nucleotides. Here, we Identify a long-distance RNA-RNA Interaction of 6 base pairs as the basis for the translational polarity. The 3' side of the complementarity region Is located in the coat-replicase intercistronic region, some 20 nucleotides before the start codon of the replicase. The 5' side encodes amino acids 31 and 32 of the coat protein. Mutations that disrupt the long-range Interaction abolish the translational coupling. Repair of basepalring by second site base substitutions restores translational coupling.


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