Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1997 Aug;41(8):1749–1754. doi: 10.1128/aac.41.8.1749

An extracellular factor regulating expression of the chromosomal aminoglycoside 2'-N-acetyltransferase of Providencia stuartii.

P N Rather 1, M M Parojcic 1, M R Paradise 1
PMCID: PMC163998  PMID: 9257754

Abstract

The chromosomal aac(2')-Ia gene in Providencia stuartii encodes a housekeeping 2'-N-acetyltransferase [AAC(2')-Ia] involved in the acetylation of peptidoglycan. In addition, the AAC(2')-Ia enzyme also acetylates and confers resistance to the clinically important aminoglycoside antibiotics gentamicin, tobramycin, and netilmicin. Expression of the aac(2')-Ia gene was found to be strongly influenced by cell density, with a sharp decrease in aac(2')-Ia mRNA accumulation as cells approached stationary phase. This decrease was mediated by the accumulation of an extracellular factor, designated AR (for acetyltransferase repressing)-factor. AR-factor was produced in both minimal and rich media and acted in a manner that was strongly dose dependent. The activity of AR-factor was also pH dependent, with optimal activity at pH 8.0 and above. Biochemical characterization of conditioned media from P. stuartii has shown that AR-factor is between 500 and 1,000 Da in molecular size and is heat stable. In addition, AR-factor was inactivated by a variety of proteases, suggesting that it may be a small peptide.

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. Bainton N. J., Bycroft B. W., Chhabra S. R., Stead P., Gledhill L., Hill P. J., Rees C. E., Winson M. K., Salmond G. P., Stewart G. S. A general role for the lux autoinducer in bacterial cell signalling: control of antibiotic biosynthesis in Erwinia. Gene. 1992 Jul 1;116(1):87–91. doi: 10.1016/0378-1119(92)90633-z. [DOI] [PubMed] [Google Scholar]
  2. Balaban N., Novick R. P. Autocrine regulation of toxin synthesis by Staphylococcus aureus. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1619–1623. doi: 10.1073/pnas.92.5.1619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cao J. G., Meighen E. A. Purification and structural identification of an autoinducer for the luminescence system of Vibrio harveyi. J Biol Chem. 1989 Dec 25;264(36):21670–21676. [PubMed] [Google Scholar]
  4. Chevereau M., Daniels P. J., Davies J., LeGoffic F. Aminoglycoside resistance in bacteria medicated by gentamicin acetyltransferase II, an enzyme modifying the 2'-amino group of aminoglycoside antibiotics. Biochemistry. 1974 Jan 29;13(3):598–603. doi: 10.1021/bi00700a030. [DOI] [PubMed] [Google Scholar]
  5. Clarke A. J. Extent of peptidoglycan O acetylation in the tribe Proteeae. J Bacteriol. 1993 Jul;175(14):4550–4553. doi: 10.1128/jb.175.14.4550-4553.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clewell D. B., Weaver K. E. Sex pheromones and plasmid transfer in Enterococcus faecalis. Plasmid. 1989 May;21(3):175–184. doi: 10.1016/0147-619x(89)90041-3. [DOI] [PubMed] [Google Scholar]
  7. Downard J., Toal D. Branched-chain fatty acids: the case for a novel form of cell-cell signalling during Myxococcus xanthus development. Mol Microbiol. 1995 Apr;16(2):171–175. doi: 10.1111/j.1365-2958.1995.tb02290.x. [DOI] [PubMed] [Google Scholar]
  8. Dunny G. M., Brown B. L., Clewell D. B. Induced cell aggregation and mating in Streptococcus faecalis: evidence for a bacterial sex pheromone. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3479–3483. doi: 10.1073/pnas.75.7.3479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eberhard A., Burlingame A. L., Eberhard C., Kenyon G. L., Nealson K. H., Oppenheimer N. J. Structural identification of autoinducer of Photobacterium fischeri luciferase. Biochemistry. 1981 Apr 28;20(9):2444–2449. doi: 10.1021/bi00512a013. [DOI] [PubMed] [Google Scholar]
  10. Eberhard A., Burlingame A. L., Eberhard C., Kenyon G. L., Nealson K. H., Oppenheimer N. J. Structural identification of autoinducer of Photobacterium fischeri luciferase. Biochemistry. 1981 Apr 28;20(9):2444–2449. doi: 10.1021/bi00512a013. [DOI] [PubMed] [Google Scholar]
  11. Fuqua C., Burbea M., Winans S. C. Activity of the Agrobacterium Ti plasmid conjugal transfer regulator TraR is inhibited by the product of the traM gene. J Bacteriol. 1995 Mar;177(5):1367–1373. doi: 10.1128/jb.177.5.1367-1373.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fuqua W. C., Winans S. C., Greenberg E. P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators. J Bacteriol. 1994 Jan;176(2):269–275. doi: 10.1128/jb.176.2.269-275.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibson M. M., Price M., Higgins C. F. Genetic characterization and molecular cloning of the tripeptide permease (tpp) genes of Salmonella typhimurium. J Bacteriol. 1984 Oct;160(1):122–130. doi: 10.1128/jb.160.1.122-130.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Grossman A. D., Losick R. Extracellular control of spore formation in Bacillus subtilis. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4369–4373. doi: 10.1073/pnas.85.12.4369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hiles I. D., Powell L. M., Higgins C. F. Peptide transport in Salmonella typhimurium: molecular cloning and characterization of the oligopeptide permease genes. Mol Gen Genet. 1987 Jan;206(1):101–109. doi: 10.1007/BF00326543. [DOI] [PubMed] [Google Scholar]
  16. Hofmeister A. E., Londoño-Vallejo A., Harry E., Stragier P., Losick R. Extracellular signal protein triggering the proteolytic activation of a developmental transcription factor in B. subtilis. Cell. 1995 Oct 20;83(2):219–226. doi: 10.1016/0092-8674(95)90163-9. [DOI] [PubMed] [Google Scholar]
  17. Huisman G. W., Kolter R. Sensing starvation: a homoserine lactone--dependent signaling pathway in Escherichia coli. Science. 1994 Jul 22;265(5171):537–539. doi: 10.1126/science.7545940. [DOI] [PubMed] [Google Scholar]
  18. Hwang I., Cook D. M., Farrand S. K. A new regulatory element modulates homoserine lactone-mediated autoinduction of Ti plasmid conjugal transfer. J Bacteriol. 1995 Jan;177(2):449–458. doi: 10.1128/jb.177.2.449-458.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Håvarstein L. S., Coomaraswamy G., Morrison D. A. An unmodified heptadecapeptide pheromone induces competence for genetic transformation in Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11140–11144. doi: 10.1073/pnas.92.24.11140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jacobs C., Joris B., Jamin M., Klarsov K., Van Beeumen J., Mengin-Lecreulx D., van Heijenoort J., Park J. T., Normark S., Frère J. M. AmpD, essential for both beta-lactamase regulation and cell wall recycling, is a novel cytosolic N-acetylmuramyl-L-alanine amidase. Mol Microbiol. 1995 Feb;15(3):553–559. doi: 10.1111/j.1365-2958.1995.tb02268.x. [DOI] [PubMed] [Google Scholar]
  21. Ji G., Beavis R. C., Novick R. P. Cell density control of staphylococcal virulence mediated by an octapeptide pheromone. Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12055–12059. doi: 10.1073/pnas.92.26.12055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Joachimiak A., Kelley R. L., Gunsalus R. P., Yanofsky C., Sigler P. B. Purification and characterization of trp aporepressor. Proc Natl Acad Sci U S A. 1983 Feb;80(3):668–672. doi: 10.1073/pnas.80.3.668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jones S., Yu B., Bainton N. J., Birdsall M., Bycroft B. W., Chhabra S. R., Cox A. J., Golby P., Reeves P. J., Stephens S. The lux autoinducer regulates the production of exoenzyme virulence determinants in Erwinia carotovora and Pseudomonas aeruginosa. EMBO J. 1993 Jun;12(6):2477–2482. doi: 10.1002/j.1460-2075.1993.tb05902.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kaiser D., Losick R. How and why bacteria talk to each other. Cell. 1993 Jun 4;73(5):873–885. doi: 10.1016/0092-8674(93)90268-u. [DOI] [PubMed] [Google Scholar]
  25. Kim S. K., Kaiser D. C-factor: a cell-cell signaling protein required for fruiting body morphogenesis of M. xanthus. Cell. 1990 Apr 6;61(1):19–26. doi: 10.1016/0092-8674(90)90211-v. [DOI] [PubMed] [Google Scholar]
  26. Kuo A., Callahan S. M., Dunlap P. V. Modulation of luminescence operon expression by N-octanoyl-L-homoserine lactone in ainS mutants of Vibrio fischeri. J Bacteriol. 1996 Feb;178(4):971–976. doi: 10.1128/jb.178.4.971-976.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kuspa A., Kroos L., Kaiser D. Intercellular signaling is required for developmental gene expression in Myxococcus xanthus. Dev Biol. 1986 Sep;117(1):267–276. doi: 10.1016/0012-1606(86)90369-6. [DOI] [PubMed] [Google Scholar]
  28. Kuspa A., Plamann L., Kaiser D. Identification of heat-stable A-factor from Myxococcus xanthus. J Bacteriol. 1992 May;174(10):3319–3326. doi: 10.1128/jb.174.10.3319-3326.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Latifi A., Winson M. K., Foglino M., Bycroft B. W., Stewart G. S., Lazdunski A., Williams P. Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PAO1. Mol Microbiol. 1995 Jul;17(2):333–343. doi: 10.1111/j.1365-2958.1995.mmi_17020333.x. [DOI] [PubMed] [Google Scholar]
  30. Macinga D. R., Parojcic M. M., Rather P. N. Identification and analysis of aarP, a transcriptional activator of the 2'-N-acetyltransferase in Providencia stuartii. J Bacteriol. 1995 Jun;177(12):3407–3413. doi: 10.1128/jb.177.12.3407-3413.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Macinga D. R., Rather P. N. aarD, a Providencia stuartii homologue of cydD: role in 2'-N-acetyltransferase expression, cell morphology and growth in the presence of an extracellular factor. Mol Microbiol. 1996 Feb;19(3):511–520. doi: 10.1046/j.1365-2958.1996.385912.x. [DOI] [PubMed] [Google Scholar]
  32. Magnuson R., Solomon J., Grossman A. D. Biochemical and genetic characterization of a competence pheromone from B. subtilis. Cell. 1994 Apr 22;77(2):207–216. doi: 10.1016/0092-8674(94)90313-1. [DOI] [PubMed] [Google Scholar]
  33. Park J. T. Turnover and recycling of the murein sacculus in oligopeptide permease-negative strains of Escherichia coli: indirect evidence for an alternative permease system and for a monolayered sacculus. J Bacteriol. 1993 Jan;175(1):7–11. doi: 10.1128/jb.175.1.7-11.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Passador L., Cook J. M., Gambello M. J., Rust L., Iglewski B. H. Expression of Pseudomonas aeruginosa virulence genes requires cell-to-cell communication. Science. 1993 May 21;260(5111):1127–1130. doi: 10.1126/science.8493556. [DOI] [PubMed] [Google Scholar]
  35. Payie K. G., Rather P. N., Clarke A. J. Contribution of gentamicin 2'-N-acetyltransferase to the O acetylation of peptidoglycan in Providencia stuartii. J Bacteriol. 1995 Aug;177(15):4303–4310. doi: 10.1128/jb.177.15.4303-4310.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pearson J. P., Gray K. M., Passador L., Tucker K. D., Eberhard A., Iglewski B. H., Greenberg E. P. Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):197–201. doi: 10.1073/pnas.91.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Piper K. R., Beck von Bodman S., Farrand S. K. Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction. Nature. 1993 Apr 1;362(6419):448–450. doi: 10.1038/362448a0. [DOI] [PubMed] [Google Scholar]
  38. Pirhonen M., Flego D., Heikinheimo R., Palva E. T. A small diffusible signal molecule is responsible for the global control of virulence and exoenzyme production in the plant pathogen Erwinia carotovora. EMBO J. 1993 Jun;12(6):2467–2476. doi: 10.1002/j.1460-2075.1993.tb05901.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rather P. N., Orosz E. Characterization of aarA, a pleiotrophic negative regulator of the 2'-N-acetyltransferase in Providencia stuartii. J Bacteriol. 1994 Aug;176(16):5140–5144. doi: 10.1128/jb.176.16.5140-5144.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rather P. N., Orosz E., Shaw K. J., Hare R., Miller G. Characterization and transcriptional regulation of the 2'-N-acetyltransferase gene from Providencia stuartii. J Bacteriol. 1993 Oct;175(20):6492–6498. doi: 10.1128/jb.175.20.6492-6498.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rather P. N., Solinsky K. A., Paradise M. R., Parojcic M. M. aarC, an essential gene involved in density-dependent regulation of the 2'-N-acetyltransferase in Providencia stuartii. J Bacteriol. 1997 Apr;179(7):2267–2273. doi: 10.1128/jb.179.7.2267-2273.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rose J. K., Yanofsky C. Interaction of the operator of the tryptophan operon with repressor. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3134–3138. doi: 10.1073/pnas.71.8.3134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Salmond G. P., Bycroft B. W., Stewart G. S., Williams P. The bacterial 'enigma': cracking the code of cell-cell communication. Mol Microbiol. 1995 May;16(4):615–624. doi: 10.1111/j.1365-2958.1995.tb02424.x. [DOI] [PubMed] [Google Scholar]
  44. Willey J., Santamaria R., Guijarro J., Geistlich M., Losick R. Extracellular complementation of a developmental mutation implicates a small sporulation protein in aerial mycelium formation by S. coelicolor. Cell. 1991 May 17;65(4):641–650. doi: 10.1016/0092-8674(91)90096-h. [DOI] [PubMed] [Google Scholar]
  45. Yamaguchi M., Mitsuhashi S., Kobayashi F., Zenda H. A 2'-N-acetylating enzyme of aminoglycosides. J Antibiot (Tokyo) 1974 Jul;27(7):507–515. doi: 10.7164/antibiotics.27.507. [DOI] [PubMed] [Google Scholar]
  46. Zhang L., Murphy P. J., Kerr A., Tate M. E. Agrobacterium conjugation and gene regulation by N-acyl-L-homoserine lactones. Nature. 1993 Apr 1;362(6419):446–448. doi: 10.1038/362446a0. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES