Analysis of the in vivo decay of the Escherichia coli dicistronic pyrF-orfF transcript: evidence for multiple degradation pathways

Journal of Molecular Biology
C M ArraianoS R Kushner

Abstract

Messenger RNA decay in Escherichia coli is slowed in pnp-7 (PNPase) rnb-500 (RNase II) rne-1(RNase E) multiple mutants. We have used Northern blots, S1 nuclease protection and primer extension analysis to map 18 endonucleolytic cleavage sites within the pyrF-orfF dicistronic transcript. Although examination of a total of 27 cleavage sites including those determined for the monocistronic trxA transcript revealed a complex pattern, the central four nucleotides within a cluster of 12 residues encompassing the cleavage sites showed a definite A/U preference. Also of interest was the processing of the dicistronic transcript to remove the downstream orfF sequence as a stable but untranslated RNA fragment. The data provide further support for the hypothesis that multiple decay pathways are involved in the decay of a single transcript. In particular, the pyrF-orfF transcript apparently can be degraded either in the 5' to 3' or the 3' to 5' direction. Our results are discussed in light of current models of mRNA decay involving polyadenylation and multiprotein decay complexes.

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Citations

Nov 20, 2001·Nucleic Acids Research·S JägerG Klug
Aug 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·R A Lease, M Belfort
Mar 25, 2006·Molecular Microbiology·Jorge M SantosCecília M Arraiano
Sep 23, 2006·FEMS Microbiology Reviews·Vladimir R Kaberdin, Udo Bläsi
Jul 28, 2010·FEMS Microbiology Reviews·Cecília M ArraianoSandra C Viegas
Jun 30, 2006·Journal of Molecular Biology·Mónica AmblarCecília M Arraiano
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Feb 4, 2014·Journal of Bacteriology·Kathrin BaumgardtElena Evguenieva-Hackenberg
Jun 17, 1999·FEMS Microbiology Reviews·R Rauhut, G Klug
Apr 1, 2007·EcoSal Plus·Sidney R Kushner

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