Abstract
The transcriptional start point for the amidase structural gene (amiE) of Pseudomonas aeruginosa has been identified, and the promoter (pE) has been shown to function constitutively, as predicted for a system regulated by transcription antitermination. Northern (RNA) analysis results show that in cells grown under inducing conditions, a major 1.3-kb amiE transcript arises from pE, and in addition, a larger transcript of approximately 5.0 kb in length has been shown to derive from the same promoter, encoding all of the genes of the operon. DNA sequencing and S1 nuclease mapping have located a transcription terminator downstream of amiE, which terminates approximately half of the pE transcripts. Previously, two RpoN-dependent promoter-like sequences (pN1 and pN2) were identified upstream of the negative regulator gene, amiC, and we have now constructed a promoter probe vector which shows weak constitutive promoter activity within this region. This promoter would be expected to provide basal levels of expression of the amiC and amiR regulatory genes to allow induction of the system.
Full Text
The Full Text of this article is available as a PDF (306.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BRAMMAR W. J., CLARKE P. H. INDUCTION AND REPRESSION OF PSEUDOMONAS AERUGINOSA AMIDASE. J Gen Microbiol. 1964 Dec;37:307–319. doi: 10.1099/00221287-37-3-307. [DOI] [PubMed] [Google Scholar]
- Brammar W. J., Charles I. G., Matfield M., Liu C. P., Drew R. E., Clarke P. H. The nucleotide sequence of the amiE gene of Pseudomonas aeruginosa. FEBS Lett. 1987 May 11;215(2):291–294. doi: 10.1016/0014-5793(87)80164-3. [DOI] [PubMed] [Google Scholar]
- Burke J. F. High-sensitivity S1 mapping with single-stranded [32P]DNA probes synthesized from bacteriophage M13mp templates. Gene. 1984 Oct;30(1-3):63–68. doi: 10.1016/0378-1119(84)90105-7. [DOI] [PubMed] [Google Scholar]
- Clarke L., Carbon J. Functional expression of cloned yeast DNA in Escherichia coli: specific complementation of argininosuccinate lyase (argH) mutations. J Mol Biol. 1978 Apr 25;120(4):517–532. doi: 10.1016/0022-2836(78)90351-0. [DOI] [PubMed] [Google Scholar]
- Clarke P. H., Drew R. E., Turberville C., Brammar W. J., Ambler R. P., Auffret A. D. Alignment of cloned amiE gene of Pseudomonas aeruginosa with the N-terminal sequence of amidase. Biosci Rep. 1981 Apr;1(4):299–307. doi: 10.1007/BF01114869. [DOI] [PubMed] [Google Scholar]
- Cousens D. J., Clarke P. H., Drew R. The amidase regulatory gene (amiR) of Pseudomonas aeruginosa. J Gen Microbiol. 1987 Aug;133(8):2041–2052. doi: 10.1099/00221287-133-8-2041. [DOI] [PubMed] [Google Scholar]
- Deretic V., Gill J. F., Chakrabarty A. M. Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa. J Bacteriol. 1987 Jan;169(1):351–358. doi: 10.1128/jb.169.1.351-358.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drew R. E., Clarke P. H., Brammar W. J. The construction in vitro of derivatives of bacteriophage lambda carrying the amidase genes of Pseudomonas aeruginosa. Mol Gen Genet. 1980 Jan;177(2):311–320. doi: 10.1007/BF00267444. [DOI] [PubMed] [Google Scholar]
- Drew R. Complementation analysis of the aliphatic amidase genes of Pseudomonas aeruginosa. J Gen Microbiol. 1984 Dec;130(12):3101–3111. doi: 10.1099/00221287-130-12-3101. [DOI] [PubMed] [Google Scholar]
- Drew R., Lowe N. Positive control of Pseudomonas aeruginosa amidase synthesis is mediated by a transcription anti-termination mechanism. J Gen Microbiol. 1989 Apr;135(4):817–823. doi: 10.1099/00221287-135-4-817. [DOI] [PubMed] [Google Scholar]
- Farin F., Clarke P. H. Positive regulation of amidase synthesis in Pseudomonas aeruginosa. J Bacteriol. 1978 Aug;135(2):379–392. doi: 10.1128/jb.135.2.379-392.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KELLY M., CLARKE P. H. An inducible amidase produced by a strain of Pseudomonas aeruginosa. J Gen Microbiol. 1962 Feb;27:305–316. doi: 10.1099/00221287-27-2-305. [DOI] [PubMed] [Google Scholar]
- Lowe N., Rice P. M., Drew R. E. Nucleotide sequence of the aliphatic amidase regulator gene (amiR) of Pseudomonas aeruginosa. FEBS Lett. 1989 Mar 27;246(1-2):39–43. doi: 10.1016/0014-5793(89)80249-2. [DOI] [PubMed] [Google Scholar]
- MacGregor C. H., Wolff J. A., Arora S. K., Phibbs P. V., Jr Cloning of a catabolite repression control (crc) gene from Pseudomonas aeruginosa, expression of the gene in Escherichia coli, and identification of the gene product in Pseudomonas aeruginosa. J Bacteriol. 1991 Nov;173(22):7204–7212. doi: 10.1128/jb.173.22.7204-7212.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pearl L., O'Hara B., Drew R., Wilson S. Crystal structure of AmiC: the controller of transcription antitermination in the amidase operon of Pseudomonas aeruginosa. EMBO J. 1994 Dec 15;13(24):5810–5817. doi: 10.1002/j.1460-2075.1994.tb06924.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smyth P. F., Clarke P. H. Catabolite repression of Pseudomonas aeruginosa amidase: the effect of carbon source on amidase synthesis. J Gen Microbiol. 1975 Sep;90(1):81–90. doi: 10.1099/00221287-90-1-81. [DOI] [PubMed] [Google Scholar]
- Wilson S. A., Wachira S. J., Drew R. E., Jones D., Pearl L. H. Antitermination of amidase expression in Pseudomonas aeruginosa is controlled by a novel cytoplasmic amide-binding protein. EMBO J. 1993 Sep;12(9):3637–3642. doi: 10.1002/j.1460-2075.1993.tb06037.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson S., Drew R. Cloning and DNA sequence of amiC, a new gene regulating expression of the Pseudomonas aeruginosa aliphatic amidase, and purification of the amiC product. J Bacteriol. 1991 Aug;173(16):4914–4921. doi: 10.1128/jb.173.16.4914-4921.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]