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Journal of Bacteriology, February 2000, p. 689-695, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Different Processing of an mRNA Species in Bacillus subtilis and Escherichia coli

Martin Persson,* Elisabeth Glatz,dagger and Blanka Rutberg

Department of Microbiology, Lund University, Sölvegatan 12, S-223 62 Lund, Sweden

Received 30 July 1999/Accepted 18 October 1999

Expression of the Bacillus subtilis glpD gene, which encodes glycerol-3-phosphate (G3P) dehydrogenase, is controlled by termination or antitermination of transcription. The untranslated leader sequence of glpD contains an inverted repeat that gives rise to a transcription terminator. In the presence of G3P, the antiterminator protein GlpP binds to glpD leader mRNA and promotes readthrough of the terminator. Certain mutations in the inverted repeat of the glpD leader result in GlpP-independent, temperature-sensitive (TS) expression of glpD. The TS phenotype is due to temperature-dependent degradation of the glpD mRNA. In the presence of GlpP, the glpD mRNA is stabilized. glpD leader-lacZ fusions were integrated into the chromosomes of B. subtilis and Escherichia coli. Determination of steady-state levels of fusion mRNA in B. subtilis showed that the stability of the fusion mRNA is determined by the glpD leader part. Comparison of steady-state levels and half-lives of glpD leader-lacZ fusion mRNA in B. subtilis and E. coli revealed significant differences. A glpD leader-lacZ fusion transcript that was unstable in B. subtilis was considerably more stable in E. coli. GlpP, which stabilizes the transcript in B. subtilis, did not affect its stability in E. coli. Primer extension analysis showed that the glpD leader-lacZ fusion transcript is processed differently in B. subtilis and in E. coli. The dominating cleavage site in E. coli was barely detectable in B. subtilis. This site was shown to be a target of E. coli RNase III.

* Corresponding author. Mailing address: Department of Microbiology, Lund University, Sölvegatan 12, S-223 62 Lund, Sweden. Phone: 46 46 2224980. Fax: 46 46 157839. E-mail: martin.persson{at}mikrbiol.lu.se.

dagger Present address: Department of Infectious Diseases and Medical Microbiology, Division of Bacteriology, Lund University, Sölvegatan 23, S-223 62 Lund, Sweden.

Journal of Bacteriology, February 2000, p. 689-695, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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