Skip to main content
RNA logoLink to RNA
. 2000 Aug;6(8):1185–1193. doi: 10.1017/s135583820000073x

RNase II removes the oligo(A) tails that destabilize the rpsO mRNA of Escherichia coli.

P E Marujo 1, E Hajnsdorf 1, J Le Derout 1, R Andrade 1, C M Arraiano 1, P Régnier 1
PMCID: PMC1369992  PMID: 10943897

Abstract

Polyadenylation controls mRNA stability in procaryotes, eucaryotes, and organelles. In bacteria, oligo(A) tails synthesized by poly(A) polymerase I are the targets of the 3'-to-5' exoribonucleases: polynucleotide phosphorylase and RNase II. Here we show that RNase II very efficiently removes the oligo(A) tails that can be used as binding sites by PNPase to start degradation of the rpsO mRNA. Both enzymes are impeded by the secondary structure of the transcription terminator at the 3' end of the mRNA. RNase II mostly generates tailless transcripts harboring 2 unpaired nt downstream of the transcription terminator hairpin, whereas PNPase releases molecules that exhibit a single-stranded stretch of 5-7 nt terminated by a tail of 3-5 As. The rpsO mRNAs whose oligo(A) tails have been removed by RNase II are more stable than oligoadenylated molecules that occur in strains deficient for RNase II. Moreover, the rpsO mRNA is stabilized when RNase II is overproduced. This modulation of mRNA stability by RNase II is only observed when poly(A) polymerase I is active. These in vivo data demonstrate that RNase II protects mRNAs ending by stable terminal hairpins, such as primary transcripts, from degradation by poly(A)-dependent ribonucleases.

Full Text

The Full Text of this article is available as a PDF (707.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arraiano C. M., Yancey S. D., Kushner S. R. Stabilization of discrete mRNA breakdown products in ams pnp rnb multiple mutants of Escherichia coli K-12. J Bacteriol. 1988 Oct;170(10):4625–4633. doi: 10.1128/jb.170.10.4625-4633.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blum E., Carpousis A. J., Higgins C. F. Polyadenylation promotes degradation of 3'-structured RNA by the Escherichia coli mRNA degradosome in vitro. J Biol Chem. 1999 Feb 12;274(7):4009–4016. doi: 10.1074/jbc.274.7.4009. [DOI] [PubMed] [Google Scholar]
  3. Carpousis A. J., Van Houwe G., Ehretsmann C., Krisch H. M. Copurification of E. coli RNAase E and PNPase: evidence for a specific association between two enzymes important in RNA processing and degradation. Cell. 1994 Mar 11;76(5):889–900. doi: 10.1016/0092-8674(94)90363-8. [DOI] [PubMed] [Google Scholar]
  4. Coburn G. A., Mackie G. A. Degradation of mRNA in Escherichia coli: an old problem with some new twists. Prog Nucleic Acid Res Mol Biol. 1999;62:55–108. doi: 10.1016/s0079-6603(08)60505-x. [DOI] [PubMed] [Google Scholar]
  5. Coburn G. A., Mackie G. A. Overexpression, purification, and properties of Escherichia coli ribonuclease II. J Biol Chem. 1996 Jan 12;271(2):1048–1053. doi: 10.1074/jbc.271.2.1048. [DOI] [PubMed] [Google Scholar]
  6. Coburn G. A., Mackie G. A. Reconstitution of the degradation of the mRNA for ribosomal protein S20 with purified enzymes. J Mol Biol. 1998 Jun 26;279(5):1061–1074. doi: 10.1006/jmbi.1998.1842. [DOI] [PubMed] [Google Scholar]
  7. Deutscher M. P., Reuven N. B. Enzymatic basis for hydrolytic versus phosphorolytic mRNA degradation in Escherichia coli and Bacillus subtilis. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3277–3280. doi: 10.1073/pnas.88.8.3277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donovan W. P., Kushner S. R. Polynucleotide phosphorylase and ribonuclease II are required for cell viability and mRNA turnover in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1986 Jan;83(1):120–124. doi: 10.1073/pnas.83.1.120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ehretsmann C. P., Carpousis A. J., Krisch H. M. Specificity of Escherichia coli endoribonuclease RNase E: in vivo and in vitro analysis of mutants in a bacteriophage T4 mRNA processing site. Genes Dev. 1992 Jan;6(1):149–159. doi: 10.1101/gad.6.1.149. [DOI] [PubMed] [Google Scholar]
  10. Grunberg-Manago M. Messenger RNA stability and its role in control of gene expression in bacteria and phages. Annu Rev Genet. 1999;33:193–227. doi: 10.1146/annurev.genet.33.1.193. [DOI] [PubMed] [Google Scholar]
  11. Hajnsdorf E., Braun F., Haugel-Nielsen J., Le Derout J., Régnier P. Multiple degradation pathways of the rpsO mRNA of Escherichia coli. RNase E interacts with the 5' and 3' extremities of the primary transcript. Biochimie. 1996;78(6):416–424. doi: 10.1016/0300-9084(96)84748-1. [DOI] [PubMed] [Google Scholar]
  12. Hajnsdorf E., Braun F., Haugel-Nielsen J., Régnier P. Polyadenylylation destabilizes the rpsO mRNA of Escherichia coli. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3973–3977. doi: 10.1073/pnas.92.9.3973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hajnsdorf E., Carpousis A. J., Régnier P. Nucleolytic inactivation and degradation of the RNase III processed pnp message encoding polynucleotide phosphorylase of Escherichia coli. J Mol Biol. 1994 Jun 17;239(4):439–454. doi: 10.1006/jmbi.1994.1387. [DOI] [PubMed] [Google Scholar]
  14. Hajnsdorf E., Régnier P. E. coli RpsO mRNA decay: RNase E processing at the beginning of the coding sequence stimulates poly(A)-dependent degradation of the mRNA. J Mol Biol. 1999 Mar 5;286(4):1033–1043. doi: 10.1006/jmbi.1999.2547. [DOI] [PubMed] [Google Scholar]
  15. Hajnsdorf E., Régnier P. Host factor Hfq of Escherichia coli stimulates elongation of poly(A) tails by poly(A) polymerase I. Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1501–1505. doi: 10.1073/pnas.040549897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hajnsdorf E., Steier O., Coscoy L., Teysset L., Régnier P. Roles of RNase E, RNase II and PNPase in the degradation of the rpsO transcripts of Escherichia coli: stabilizing function of RNase II and evidence for efficient degradation in an ams pnp rnb mutant. EMBO J. 1994 Jul 15;13(14):3368–3377. doi: 10.1002/j.1460-2075.1994.tb06639.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Haugel-Nielsen J., Hajnsdorf E., Regnier P. The rpsO mRNA of Escherichia coli is polyadenylated at multiple sites resulting from endonucleolytic processing and exonucleolytic degradation. EMBO J. 1996 Jun 17;15(12):3144–3152. [PMC free article] [PubMed] [Google Scholar]
  18. Huang H., Liao J., Cohen S. N. Poly(A)- and poly(U)-specific RNA 3' tail shortening by E. coli ribonuclease E. Nature. 1998 Jan 1;391(6662):99–102. doi: 10.1038/34219. [DOI] [PubMed] [Google Scholar]
  19. Kalapos M. P., Cao G. J., Kushner S. R., Sarkar N. Identification of a second poly(A) polymerase in Escherichia coli. Biochem Biophys Res Commun. 1994 Jan 28;198(2):459–465. doi: 10.1006/bbrc.1994.1067. [DOI] [PubMed] [Google Scholar]
  20. Lapham J., Yu Y. T., Shu M. D., Steitz J. A., Crothers D. M. The position of site-directed cleavage of RNA using RNase H and 2'-O-methyl oligonucleotides is dependent on the enzyme source. RNA. 1997 Sep;3(9):950–951. [PMC free article] [PubMed] [Google Scholar]
  21. Li Z., Deutscher M. P. The tRNA processing enzyme RNase T is essential for maturation of 5S RNA. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6883–6886. doi: 10.1073/pnas.92.15.6883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Li Z., Pandit S., Deutscher M. P. 3' exoribonucleolytic trimming is a common feature of the maturation of small, stable RNAs in Escherichia coli. Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2856–2861. doi: 10.1073/pnas.95.6.2856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Li Z., Pandit S., Deutscher M. P. Maturation of 23S ribosomal RNA requires the exoribonuclease RNase T. RNA. 1999 Jan;5(1):139–146. doi: 10.1017/s1355838299981669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McLaren R. S., Newbury S. F., Dance G. S., Causton H. C., Higgins C. F. mRNA degradation by processive 3'-5' exoribonucleases in vitro and the implications for prokaryotic mRNA decay in vivo. J Mol Biol. 1991 Sep 5;221(1):81–95. [PubMed] [Google Scholar]
  25. Miczak A., Kaberdin V. R., Wei C. L., Lin-Chao S. Proteins associated with RNase E in a multicomponent ribonucleolytic complex. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3865–3869. doi: 10.1073/pnas.93.9.3865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mohanty B. K., Kushner S. R. Analysis of the function of Escherichia coli poly(A) polymerase I in RNA metabolism. Mol Microbiol. 1999 Dec;34(5):1094–1108. doi: 10.1046/j.1365-2958.1999.01673.x. [DOI] [PubMed] [Google Scholar]
  27. Mohanty B. K., Kushner S. R. Residual polyadenylation in poly(A) polymerase I (pcnB ) mutants of Escherichia coli does not result from the activity encoded by the f310 gene. Mol Microbiol. 1999 Dec;34(5):1109–1119. doi: 10.1046/j.1365-2958.1999.01674.x. [DOI] [PubMed] [Google Scholar]
  28. Mudd E. A., Krisch H. M., Higgins C. F. RNase E, an endoribonuclease, has a general role in the chemical decay of Escherichia coli mRNA: evidence that rne and ams are the same genetic locus. Mol Microbiol. 1990 Dec;4(12):2127–2135. doi: 10.1111/j.1365-2958.1990.tb00574.x. [DOI] [PubMed] [Google Scholar]
  29. O'Hara E. B., Chekanova J. A., Ingle C. A., Kushner Z. R., Peters E., Kushner S. R. Polyadenylylation helps regulate mRNA decay in Escherichia coli. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1807–1811. doi: 10.1073/pnas.92.6.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pepe C. M., Maslesa-Galić S., Simons R. W. Decay of the IS10 antisense RNA by 3' exoribonucleases: evidence that RNase II stabilizes RNA-OUT against PNPase attack. Mol Microbiol. 1994 Sep;13(6):1133–1142. doi: 10.1111/j.1365-2958.1994.tb00504.x. [DOI] [PubMed] [Google Scholar]
  31. Py B., Higgins C. F., Krisch H. M., Carpousis A. J. A DEAD-box RNA helicase in the Escherichia coli RNA degradosome. Nature. 1996 May 9;381(6578):169–172. doi: 10.1038/381169a0. [DOI] [PubMed] [Google Scholar]
  32. Régnier P., Arraiano C. M. Degradation of mRNA in bacteria: emergence of ubiquitous features. Bioessays. 2000 Mar;22(3):235–244. doi: 10.1002/(SICI)1521-1878(200003)22:3<235::AID-BIES5>3.0.CO;2-2. [DOI] [PubMed] [Google Scholar]
  33. Régnier P., Grunberg-Manago M. RNase III cleavages in non-coding leaders of Escherichia coli transcripts control mRNA stability and genetic expression. Biochimie. 1990 Nov;72(11):825–834. doi: 10.1016/0300-9084(90)90192-j. [DOI] [PubMed] [Google Scholar]
  34. Régnier P., Hajnsdorf E. Decay of mRNA encoding ribosomal protein S15 of Escherichia coli is initiated by an RNase E-dependent endonucleolytic cleavage that removes the 3' stabilizing stem and loop structure. J Mol Biol. 1991 Jan 20;217(2):283–292. doi: 10.1016/0022-2836(91)90542-e. [DOI] [PubMed] [Google Scholar]
  35. Régnier P., Portier C. Initiation, attenuation and RNase III processing of transcripts from the Escherichia coli operon encoding ribosomal protein S15 and polynucleotide phosphorylase. J Mol Biol. 1986 Jan 5;187(1):23–32. doi: 10.1016/0022-2836(86)90403-1. [DOI] [PubMed] [Google Scholar]
  36. Sarkar N. Polyadenylation of mRNA in prokaryotes. Annu Rev Biochem. 1997;66:173–197. doi: 10.1146/annurev.biochem.66.1.173. [DOI] [PubMed] [Google Scholar]
  37. Singer M. F., Tolbert G. Purification and properties of a potassium-activated phosphodiesterase (RNAase II) from Escherichia coli. Biochemistry. 1965 Jul;4(7):1319–1330. doi: 10.1021/bi00883a016. [DOI] [PubMed] [Google Scholar]
  38. Vanzo N. F., Li Y. S., Py B., Blum E., Higgins C. F., Raynal L. C., Krisch H. M., Carpousis A. J. Ribonuclease E organizes the protein interactions in the Escherichia coli RNA degradosome. Genes Dev. 1998 Sep 1;12(17):2770–2781. doi: 10.1101/gad.12.17.2770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Xu F., Cohen S. N. RNA degradation in Escherichia coli regulated by 3' adenylation and 5' phosphorylation. Nature. 1995 Mar 9;374(6518):180–183. doi: 10.1038/374180a0. [DOI] [PubMed] [Google Scholar]
  40. Xu F., Lin-Chao S., Cohen S. N. The Escherichia coli pcnB gene promotes adenylylation of antisense RNAI of ColE1-type plasmids in vivo and degradation of RNAI decay intermediates. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6756–6760. doi: 10.1073/pnas.90.14.6756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zilhão R., Cairrão F., Régnier P., Arraiano C. M. PNPase modulates RNase II expression in Escherichia coli: implications for mRNA decay and cell metabolism. Mol Microbiol. 1996 Jun;20(5):1033–1042. doi: 10.1111/j.1365-2958.1996.tb02544.x. [DOI] [PubMed] [Google Scholar]
  42. Zilhão R., Camelo L., Arraiano C. M. DNA sequencing and expression of the gene rnb encoding Escherichia coli ribonuclease II. Mol Microbiol. 1993 Apr;8(1):43–51. doi: 10.1111/j.1365-2958.1993.tb01201.x. [DOI] [PubMed] [Google Scholar]

Articles from RNA are provided here courtesy of The RNA Society

RESOURCES