Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 May;179(9):3043–3046. doi: 10.1128/jb.179.9.3043-3046.1997

Identification of greA encoding a transcriptional elongation factor as a member of the carA-orf-carB-greA operon in Pseudomonas aeruginosa PAO1.

C D Lu 1, D H Kwon 1, A T Abdelal 1
PMCID: PMC179072  PMID: 9139926

Abstract

A homolog of the transcriptional elongation factor, GreA, was identified in Pseudomonas aeruginosa PAO1. The deduced amino acid sequence for GreA from this organism exhibits 65.2% identity to its counterpart in Escherichia coli K-12. The nucleotide sequence of greA from P. aeruginosa overlaps by four bases the 3' terminus of carB which encodes the large subunit of carbamoylphosphate synthetase. S1 nuclease experiments showed that level of the greA transcript is elevated approximately 10-fold under conditions of pyrimidine limitation, consistent with the conclusion that transcription is initiated from the previously identified pyrimidine-sensitive promoter upstream of the carA-orf-carB-greA operon. Transcriptional fusion experiments showed the presence of an additional weak promoter within the carB sequence. A greA insertional mutant of Pseudomonas aerugionsa was constructed by gene replacement. The mutant derivative grew well in rich medium but did not grow in minimal medium supplemented by arginine and nucleosides. The greA phenotype was suppressed by secondary mutations at a relatively high rate, consistent with the notion of an important physiological role for GreA.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Altmann C. R., Solow-Cordero D. E., Chamberlin M. J. RNA cleavage and chain elongation by Escherichia coli DNA-dependent RNA polymerase in a binary enzyme.RNA complex. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3784–3788. doi: 10.1073/pnas.91.9.3784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borukhov S., Polyakov A., Nikiforov V., Goldfarb A. GreA protein: a transcription elongation factor from Escherichia coli. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8899–8902. doi: 10.1073/pnas.89.19.8899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borukhov S., Sagitov V., Goldfarb A. Transcript cleavage factors from E. coli. Cell. 1993 Feb 12;72(3):459–466. doi: 10.1016/0092-8674(93)90121-6. [DOI] [PubMed] [Google Scholar]
  4. Erie D. A., Hajiseyedjavadi O., Young M. C., von Hippel P. H. Multiple RNA polymerase conformations and GreA: control of the fidelity of transcription. Science. 1993 Nov 5;262(5135):867–873. doi: 10.1126/science.8235608. [DOI] [PubMed] [Google Scholar]
  5. Farinha M. A., Kropinski A. M. Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters. J Bacteriol. 1990 Jun;172(6):3496–3499. doi: 10.1128/jb.172.6.3496-3499.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gambello M. J., Iglewski B. H. Cloning and characterization of the Pseudomonas aeruginosa lasR gene, a transcriptional activator of elastase expression. J Bacteriol. 1991 May;173(9):3000–3009. doi: 10.1128/jb.173.9.3000-3009.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haas D., Holloway B. W., Schamböck A., Leisinger T. The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet. 1977 Jul 7;154(1):7–22. doi: 10.1007/BF00265571. [DOI] [PubMed] [Google Scholar]
  8. Kwon D. H., Lu C. D., Walthall D. A., Brown T. M., Houghton J. E., Abdelal A. T. Structure and regulation of the carAB operon in Pseudomonas aeruginosa and Pseudomonas stutzeri: no untranslated region exists. J Bacteriol. 1994 May;176(9):2532–2542. doi: 10.1128/jb.176.9.2532-2542.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lu C. D., Kilstrup M., Neuhard J., Abdelal A. Pyrimidine regulation of tandem promoters for carAB in Salmonella typhimurium. J Bacteriol. 1989 Oct;171(10):5436–5442. doi: 10.1128/jb.171.10.5436-5442.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mierendorf R. C., Pfeffer D. Direct sequencing of denatured plasmid DNA. Methods Enzymol. 1987;152:556–562. doi: 10.1016/0076-6879(87)52061-4. [DOI] [PubMed] [Google Scholar]
  11. Nudler E., Kashlev M., Nikiforov V., Goldfarb A. Coupling between transcription termination and RNA polymerase inchworming. Cell. 1995 May 5;81(3):351–357. doi: 10.1016/0092-8674(95)90388-7. [DOI] [PubMed] [Google Scholar]
  12. Orlova M., Newlands J., Das A., Goldfarb A., Borukhov S. Intrinsic transcript cleavage activity of RNA polymerase. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4596–4600. doi: 10.1073/pnas.92.10.4596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roland K. L., Powell F. E., Turnbough C. L., Jr Role of translation and attenuation in the control of pyrBI operon expression in Escherichia coli K-12. J Bacteriol. 1985 Sep;163(3):991–999. doi: 10.1128/jb.163.3.991-999.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schweizer H. D. Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. Biotechniques. 1993 Nov;15(5):831–834. [PubMed] [Google Scholar]
  15. Sparkowski J., Das A. Location of a new gene, greA, on the Escherichia coli chromosome. J Bacteriol. 1991 Sep;173(17):5256–5257. doi: 10.1128/jb.173.17.5256-5257.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sparkowski J., Das A. The nucleotide sequence of greA, a suppressor gene that restores growth of an Escherichia coli RNA polymerase mutant at high temperature. Nucleic Acids Res. 1990 Nov 11;18(21):6443–6443. doi: 10.1093/nar/18.21.6443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stalon V., Ramos F., Piérard A., Wiame J. M. The occurrence of a catabolic and an anabolic ornithine carbamoyltransferase in Pseudomonas. Biochim Biophys Acta. 1967 May 16;139(1):91–97. doi: 10.1016/0005-2744(67)90115-5. [DOI] [PubMed] [Google Scholar]
  18. Stibitz S., Black W., Falkow S. The construction of a cloning vector designed for gene replacement in Bordetella pertussis. Gene. 1986;50(1-3):133–140. doi: 10.1016/0378-1119(86)90318-5. [DOI] [PubMed] [Google Scholar]
  19. Surratt C. K., Milan S. C., Chamberlin M. J. Spontaneous cleavage of RNA in ternary complexes of Escherichia coli RNA polymerase and its significance for the mechanism of transcription. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7983–7987. doi: 10.1073/pnas.88.18.7983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES