Abstract
The maturation of 5S RNA in Escherichia coli is poorly understood. Although it is known that large precursors of 5S RNA accumulate in mutant cells lacking the endoribonuclease-RNase E, almost nothing is known about how the mature 5' and 3' termini of these molecules are generated. We have examined 5S RNA maturation in wild-type and single- or multiple-exoribonuclease-deficient cells by Northern blot and primer-extension analysis. Our results indicate that no mature 5S RNA is made in RNase T-deficient strains. Rather, 5S RNA precursors containing predominantly 2 extra nucleotides at the 3' end accumulate. Apparently, these 5S RNAs are functional inasmuch as mutant cells are viable, growing only slightly slower than wild type. Purified RNase T can remove the extra 3' residues, showing that it is directly involved in the trimming reaction. In contrast, mutations affecting other 3' exoribonucleases have no effect on 5S RNA maturation. Approximately 90% of the 5S RNAs in both wild-type and RNase T- cells contain mature 5' termini, indicating that 5' processing is independent of RNase T action. These data identify the enzyme responsible for generating the mature 3' terminus of 5S RNA molecules and also demonstrate that a completely processed 5S RNA molecule is not essential for cell survival.
Full text
PDF



Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Boorstein W. R., Craig E. A. Primer extension analysis of RNA. Methods Enzymol. 1989;180:347–369. doi: 10.1016/0076-6879(89)80111-9. [DOI] [PubMed] [Google Scholar]
- Brosius J., Dull T. J., Sleeter D. D., Noller H. F. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol. 1981 May 15;148(2):107–127. doi: 10.1016/0022-2836(81)90508-8. [DOI] [PubMed] [Google Scholar]
- Dahlberg A. E., Dahlberg J. E., Lund E., Tokimatsu H., Rabson A. B., Calvert P. C., Reynolds F., Zahalak M. Processing of the 5' end of Escherichia coli 16S ribosomal RNA. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3598–3602. doi: 10.1073/pnas.75.8.3598. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deutscher M. P., Hilderman R. H. Isolation and partial characterization of Escherichia coli mutants with low levels of transfer ribonucleic acid nucleotidyltransferase. J Bacteriol. 1974 May;118(2):621–627. doi: 10.1128/jb.118.2.621-627.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deutscher M. P., Marlor C. W. Purification and characterization of Escherichia coli RNase T. J Biol Chem. 1985 Jun 10;260(11):7067–7071. [PubMed] [Google Scholar]
- Deutscher M. P., Marlor C. W., Zaniewski R. Ribonuclease T: new exoribonuclease possibly involved in end-turnover of tRNA. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4290–4293. doi: 10.1073/pnas.81.14.4290. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deutscher M. P. Promiscuous exoribonucleases of Escherichia coli. J Bacteriol. 1993 Aug;175(15):4577–4583. doi: 10.1128/jb.175.15.4577-4583.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deutscher M. P. Ribonuclease multiplicity, diversity, and complexity. J Biol Chem. 1993 Jun 25;268(18):13011–13014. [PubMed] [Google Scholar]
- Feunteun J., Jordan B. R., Monier R. Study of the maturation of 5 s RNA precursors in Escherichia coli. J Mol Biol. 1972 Oct 14;70(3):465–474. doi: 10.1016/0022-2836(72)90553-0. [DOI] [PubMed] [Google Scholar]
- Hayes F., Vasseur M. Processing of the 17-S Escherichia coli precursor RNA in the 27-S pre-ribosomal particle. Eur J Biochem. 1976 Jan 15;61(2):433–442. doi: 10.1111/j.1432-1033.1976.tb10037.x. [DOI] [PubMed] [Google Scholar]
- Kelly K. O., Deutscher M. P. The presence of only one of five exoribonucleases is sufficient to support the growth of Escherichia coli. J Bacteriol. 1992 Oct;174(20):6682–6684. doi: 10.1128/jb.174.20.6682-6684.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li Z., Deutscher M. P. The role of individual exoribonucleases in processing at the 3' end of Escherichia coli tRNA precursors. J Biol Chem. 1994 Feb 25;269(8):6064–6071. [PubMed] [Google Scholar]
- Meyhack B., Pace B., Pace N. R. Involvement of precursor-specific segments in the in vitro maturation of Bacillus subtilis precursor 5S ribosomal RNA. Biochemistry. 1977 Nov 15;16(23):5009–5015. doi: 10.1021/bi00642a011. [DOI] [PubMed] [Google Scholar]
- Meyhack B., Pace N. R. Involvement of the mature domain in the in vitro maturation of Bacillus subtilis precursor 5S ribosomal RNA. Biochemistry. 1978 Dec 26;17(26):5804–5810. doi: 10.1021/bi00619a030. [DOI] [PubMed] [Google Scholar]
- Misra T. K., Apirion D. RNase E, an RNA processing enzyme from Escherichia coli. J Biol Chem. 1979 Nov 10;254(21):11154–11159. [PubMed] [Google Scholar]
- Padmanabha K. P., Deutscher M. P. RNase T affects Escherichia coli growth and recovery from metabolic stress. J Bacteriol. 1991 Feb;173(4):1376–1381. doi: 10.1128/jb.173.4.1376-1381.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ray B. K., Singh B., Roy M. K., Apirion D. Ribonuclease E is involved in the processing of 5-S rRNA from a number of rRNA transcription units. Eur J Biochem. 1982 Jul;125(2):283–289. doi: 10.1111/j.1432-1033.1982.tb06680.x. [DOI] [PubMed] [Google Scholar]
- Reuven N. B., Deutscher M. P. Multiple exoribonucleases are required for the 3' processing of Escherichia coli tRNA precursors in vivo. FASEB J. 1993 Jan;7(1):143–148. doi: 10.1096/fasebj.7.1.8422961. [DOI] [PubMed] [Google Scholar]
- Roy M. K., Singh B., Ray B. K., Apirion D. Maturation of 5-S rRNA: ribonuclease E cleavages and their dependence on precursor sequences. Eur J Biochem. 1983 Mar 1;131(1):119–127. doi: 10.1111/j.1432-1033.1983.tb07238.x. [DOI] [PubMed] [Google Scholar]
- Sirdeshmukh R., Schlessinger D. Why is processing of 23 S ribosomal RNA in Escherichia coli not obligate for its function? J Mol Biol. 1985 Dec 5;186(3):669–672. doi: 10.1016/0022-2836(85)90139-1. [DOI] [PubMed] [Google Scholar]
- Sogin M. L., Pace B., Pace N. R. Partial purification and properties of a ribosomal RNA maturation endonuclease from Bacillus subtilis. J Biol Chem. 1977 Feb 25;252(4):1350–1357. [PubMed] [Google Scholar]
- Srivastava A. K., Schlessinger D. Coregulation of processing and translation: mature 5' termini of Escherichia coli 23S ribosomal RNA form in polysomes. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7144–7148. doi: 10.1073/pnas.85.19.7144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Srivastava A. K., Schlessinger D. Mechanism and regulation of bacterial ribosomal RNA processing. Annu Rev Microbiol. 1990;44:105–129. doi: 10.1146/annurev.mi.44.100190.000541. [DOI] [PubMed] [Google Scholar]
- Srivastava A. K., Schlessinger D. Preparation of extracts and assay of ribosomal RNA maturation in Escherichia coli. Methods Enzymol. 1990;181:355–366. doi: 10.1016/0076-6879(90)81135-h. [DOI] [PubMed] [Google Scholar]