Polyadenylylation Helps Regulate mRNA Decay in Escherichia coli

As part of our genetic analysis of mRNA decay in Escherichia coli K-12, we examined the effect of the pcnB gene [encoding poly(A) polymerase I] on message stability. Eliminating poly(A) polymerase I (ΔpcnB) dramatically stabilized the lpp, ompA, and trxA transcripts. The half-lives of individual mRN...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1995-03, Vol.92 (6), p.1807-1811
Main Authors: O'Hara, Eileen B., Chekanova, Julia A., Ingle, Caroline A., Kushner, Ze'Eva R., Peters, Erica, Kushner, Sidney R.
Format: Article
Language:English
Subjects:
Bacteria
Blotting, Northern
DNA
Enzymes
Escherichia coli
Escherichia coli - metabolism
Gels
Gene Expression
Genes, Bacterial
Genetic hybridization
Genetics
Half lives
Half-Life
Homeostasis
Messenger RNA
Mutagenesis
Poly A - analysis
Polynucleotide Adenylyltransferase - biosynthesis
Polynucleotide Adenylyltransferase - genetics
Polynucleotide Adenylyltransferase - metabolism
Polynucleotides
Ribonucleic acid
RNA
RNA stability
RNA, Bacterial - genetics
RNA, Bacterial - metabolism
RNA, Messenger - metabolism
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Summary:As part of our genetic analysis of mRNA decay in Escherichia coli K-12, we examined the effect of the pcnB gene [encoding poly(A) polymerase I] on message stability. Eliminating poly(A) polymerase I (ΔpcnB) dramatically stabilized the lpp, ompA, and trxA transcripts. The half-lives of individual mRNAs were increased in both a ΔpcnB single mutant and a ΔpcnB pnp-7 rnb-500 rne-1 multiple mutant. We also found mRNA decay intermediates in ΔpcnB mutants that were not detected in control strains. By end-labeling total E. coli RNA with [32P]pCp and T4 RNA ligase and then digesting the RNA with RNase A and T1, we showed that many RNAs in a wild-type strain contained poly(A) tails ranging from 10nt to >50 nt long. When polynucleotide phosphorylase, RNase II, and RNase E were absent, the length (>100 nt) and number (10- to 20-fold) of the poly(A) tails increased. After transcription initiation was stopped with rifampicin, polyadenylylation apparently continued. Deleting the structural gene for poly(A) polymerase I (pcnB) reduced the amount of 3'-terminal poly(A) sequences by >90%. We propose a model for the role of polyadenylylation in mRNA decay.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.92.6.1807