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. 1997 Dec;179(23):7379–7385. doi: 10.1128/jb.179.23.7379-7385.1997

Bacillus subtilis RNase III gene: cloning, function of the gene in Escherichia coli, and construction of Bacillus subtilis strains with altered rnc loci.

W Wang 1, D H Bechhofer 1
PMCID: PMC179688  PMID: 9393702

Abstract

The rnc gene of Bacillus subtilis, which has 36% amino acid identity with the gene that encodes Escherichia coli RNase III endonuclease, was cloned in E. coli and shown by functional assays to encode B. subtilis RNase III (Bs-RNase III). The cloned B. subtilis rnc gene could complement an E. coli rnc strain that is deficient in rRNA processing, suggesting that Bs-RNase III is involved in rRNA processing in B. subtilis. Attempts to construct a B. subtilis rnc null mutant were unsuccessful, but a strain was constructed in which only a carboxy-terminal truncated version of Bs-RNase III was expressed. The truncated Bs-RNase III showed virtually no activity in vitro but was active in vivo. Analysis of expression of a copy of the rnc gene integrated at the amy locus and transcribed from a p(spac) promoter suggested that expression of the B. subtilis rnc is under regulatory control.

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Selected References

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  1. Apirion D., Neil J., Watson N. Consequences of losing ribonuclease III on the Escherichia coli cell. Mol Gen Genet. 1976 Mar 22;144(2):185–190. doi: 10.1007/BF02428107. [DOI] [PubMed] [Google Scholar]
  2. Babitzke P., Granger L., Olszewski J., Kushner S. R. Analysis of mRNA decay and rRNA processing in Escherichia coli multiple mutants carrying a deletion in RNase III. J Bacteriol. 1993 Jan;175(1):229–239. doi: 10.1128/jb.175.1.229-239.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bardwell J. C., Régnier P., Chen S. M., Nakamura Y., Grunberg-Manago M., Court D. L. Autoregulation of RNase III operon by mRNA processing. EMBO J. 1989 Nov;8(11):3401–3407. doi: 10.1002/j.1460-2075.1989.tb08504.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bram R. J., Young R. A., Steitz J. A. The ribonuclease III site flanking 23S sequences in the 30S ribosomal precursor RNA of E. coli. Cell. 1980 Feb;19(2):393–401. doi: 10.1016/0092-8674(80)90513-9. [DOI] [PubMed] [Google Scholar]
  5. Brosius J. Compilation of superlinker vectors. Methods Enzymol. 1992;216:469–483. doi: 10.1016/0076-6879(92)16043-j. [DOI] [PubMed] [Google Scholar]
  6. DiMari J. F., Bechhofer D. H. Initiation of mRNA decay in Bacillus subtilis. Mol Microbiol. 1993 Mar;7(5):705–717. doi: 10.1111/j.1365-2958.1993.tb01161.x. [DOI] [PubMed] [Google Scholar]
  7. Dubnau D., Davidoff-Abelson R. Fate of transforming DNA following uptake by competent Bacillus subtilis. I. Formation and properties of the donor-recipient complex. J Mol Biol. 1971 Mar 14;56(2):209–221. doi: 10.1016/0022-2836(71)90460-8. [DOI] [PubMed] [Google Scholar]
  8. Grant S. G., Jessee J., Bloom F. R., Hanahan D. Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4645–4649. doi: 10.1073/pnas.87.12.4645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henner D. J. Inducible expression of regulatory genes in Bacillus subtilis. Methods Enzymol. 1990;185:223–228. doi: 10.1016/0076-6879(90)85022-g. [DOI] [PubMed] [Google Scholar]
  10. Ireton K., Rudner D. Z., Siranosian K. J., Grossman A. D. Integration of multiple developmental signals in Bacillus subtilis through the Spo0A transcription factor. Genes Dev. 1993 Feb;7(2):283–294. doi: 10.1101/gad.7.2.283. [DOI] [PubMed] [Google Scholar]
  11. Kameyama L., Fernandez L., Court D. L., Guarneros G. RNaselll activation of bacteriophage lambda N synthesis. Mol Microbiol. 1991 Dec;5(12):2953–2963. doi: 10.1111/j.1365-2958.1991.tb01855.x. [DOI] [PubMed] [Google Scholar]
  12. Kuwano M., Ono M., Endo H., Hori K., Nakamura K., Hirota Y., Ohnishi Y. Gene affecting longevity of messenger RNA: a mutant of Escherichia coli with altered mRNA stability. Mol Gen Genet. 1977 Sep 9;154(3):279–285. doi: 10.1007/BF00571283. [DOI] [PubMed] [Google Scholar]
  13. Loughney K., Lund E., Dahlberg J. E. Ribosomal RNA precursors of Bacillus subtilis. Nucleic Acids Res. 1983 Oct 11;11(19):6709–6721. doi: 10.1093/nar/11.19.6709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mitra S., Bechhofer D. H. Substrate specificity of an RNase III-like activity from Bacillus subtilis. J Biol Chem. 1994 Dec 16;269(50):31450–31456. [PubMed] [Google Scholar]
  15. Mitra S., Hue K., Bechhofer D. H. In vitro processing activity of Bacillus subtilis polynucleotide phosphorylase. Mol Microbiol. 1996 Jan;19(2):329–342. doi: 10.1046/j.1365-2958.1996.378906.x. [DOI] [PubMed] [Google Scholar]
  16. Nikolaev N., Silengo L., Schlessinger D. Synthesis of a large precursor to ribosomal RNA in a mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3361–3365. doi: 10.1073/pnas.70.12.3361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oguro A., Kakeshita H., Takamatsu H., Nakamura K., Yamane K. The effect of Srb, a homologue of the mammalian SRP receptor alpha-subunit, on Bacillus subtilis growth and protein translocation. Gene. 1996 Jun 12;172(1):17–24. doi: 10.1016/0378-1119(96)00181-3. [DOI] [PubMed] [Google Scholar]
  18. Panganiban A. T., Whiteley H. R. Bacillus subtilis RNAase III cleavage sites in phage SP82 early mRNA. Cell. 1983 Jul;33(3):907–913. doi: 10.1016/0092-8674(83)90033-8. [DOI] [PubMed] [Google Scholar]
  19. Panganiban A. T., Whiteley H. R. Purification and properties of a new bacillus subtilis RNA processing enzyme. Cleavage of phage SP82 mRNA and Bacillus subtilis precursor rRNA. J Biol Chem. 1983 Oct 25;258(20):12487–12493. [PubMed] [Google Scholar]
  20. Portier C., Dondon L., Grunberg-Manago M., Régnier P. The first step in the functional inactivation of the Escherichia coli polynucleotide phosphorylase messenger is a ribonuclease III processing at the 5' end. EMBO J. 1987 Jul;6(7):2165–2170. doi: 10.1002/j.1460-2075.1987.tb02484.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Régnier P., Grunberg-Manago M. Cleavage by RNase III in the transcripts of the met Y-nus-A-infB operon of Escherichia coli releases the tRNA and initiates the decay of the downstream mRNA. J Mol Biol. 1989 Nov 20;210(2):293–302. doi: 10.1016/0022-2836(89)90331-8. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Wilson H. R., Kameyama L., Zhou J. G., Guarneros G., Court D. L. Translational repression by a transcriptional elongation factor. Genes Dev. 1997 Sep 1;11(17):2204–2213. doi: 10.1101/gad.11.17.2204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Young R. A., Steitz J. A. Complementary sequences 1700 nucleotides apart form a ribonuclease III cleavage site in Escherichia coli ribosomal precursor RNA. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3593–3597. doi: 10.1073/pnas.75.8.3593. [DOI] [PMC free article] [PubMed] [Google Scholar]

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