PMID:7534403
| Citation |
O'Hara, EB, Chekanova, JA, Ingle, CA, Kushner, ZR, Peters, E and Kushner, SR (1995) Polyadenylylation helps regulate mRNA decay in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 92:1807-11 |
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| Abstract |
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 (delta pcnB) dramatically stabilized the lpp, ompA, and trxA transcripts. The half-lives of individual mRNAs were increased in both a delta pcnB single mutant and a delta pcnB pnp-7 rnb-500 rne-1 multiple mutant. We also found mRNA decay intermediates in delta pcnB 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 10 nt 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. |
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| Keywords |
Blotting, Northern; Escherichia coli/metabolism; Gene Expression; Genes, Bacterial; Half-Life; Homeostasis; Mutagenesis; Poly A/analysis; Polynucleotide Adenylyltransferase/biosynthesis; Polynucleotide Adenylyltransferase/genetics; Polynucleotide Adenylyltransferase/metabolism; RNA, Bacterial/genetics; RNA, Bacterial/metabolism; RNA, Messenger/metabolism |
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