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Programmed cell death in bacteria: translational repression by mRNA end-pairing

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The hok/sok and pnd systems of plasmids R1 and R483 mediate plasmid maintenance by killing plasmid-free cells. Translation of the exceptionally stable hok and pnd mRNAs is repressed by unstable antisense RNAs. The different stabilities of the killer mRNAs and their cognate repressors explain the onset of translation in plasmid-free cells. The full-length hok and pnd mRNAs are inert with respect to translation and antisense RNA binding. We have previously shown that the mRNAs contain two negative translational control elements. Thus, the mRNAs contain upstream anti-Shine–Dalgarno elements that repress translation by shielding the Shine–Dalgarno ele-ments. The mRNAs also contain fold-back-inhibition elements (fbi ) at their 3′ ends that are required to maintain the inert mRNA configuration. Using genetic complementation, we show that the 3′fbi elements pair with the very 5′ ends of the mRNAs. This pairing sets the low rate of 3′ exonucleolytical processing, which is required for the accumulation of an activatable pool of mRNA. Unexpectedly, the hok and pnd mRNAs were found to contain translational activators at their 5′ ends (termed tac ). Thus, the fbi elements inhibit translation of the full-length mRNAs by sequestration of the tac elements. The fbi elements are removed by 3′ exonucleolytical processing. Mutational ana-lyses indicate that the 3′ processing triggers refolding of the mRNA 5′ ends into translatable configurations in which the 5′tac elements base pair with the anti-Shine–Dalgarno sequences.
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Document Type: Research Article

Affiliations: Department of Molecular Biology, Odense University, DK-5230, Odense M, Denmark.

Publication date: September 1, 1996

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