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. 1997 Apr;179(7):2267–2273. doi: 10.1128/jb.179.7.2267-2273.1997

aarC, an essential gene involved in density-dependent regulation of the 2'-N-acetyltransferase in Providencia stuartii.

P N Rather 1, K A Solinsky 1, M R Paradise 1, M M Parojcic 1
PMCID: PMC178963  PMID: 9079912

Abstract

The 2'-N-acetyltransferase [AAC(2')-Ia] in Providencia stuartii has a dual function where it is involved in the acetylation of peptidoglycan and certain aminoglycosides. A search for negative regulators of the aac(2')-Ia gene has resulted in the identification of aarC. A missense allele (aarC1) resulted in an 8.9-fold increase in beta-galactosidase accumulation from an aac(2')-lacZ transcriptional fusion. Northern blot analysis demonstrated an increase in aac(2')-Ia mRNA accumulation that was specific to cells at high density. In addition, the aarC1 allele also resulted in a substantial increase in the expression of aarP, a transcriptional activator of the aac(2')-Ia gene. The wild-type aarC gene was isolated by complementation and encodes a predicted protein of 365 amino acids with a molecular mass of 39,815 Da. The predicted AarC protein exhibited 88% amino acid homology to the previously identified GcpE protein of Escherichia coli and 86% homology to a gene product from Haemophilus influenzae. The E. coli gcpE gene was able to functionally complement the aarC1 allele in P. stuartii. The aarC1 allele was identified as a T to G transversion that resulted in a valine to glycine substitution at position 136 in the AarC protein. The aarC gene appears to be essential for cell viability as construction of a disrupted copy (aarC::lacZ) was possible only in cells that carried an episomal copy of aarC or gcpE.

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Selected References

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  1. Baker J., Franklin D. B., Parker J. Sequence and characterization of the gcpE gene of Escherichia coli. FEMS Microbiol Lett. 1992 Jul 1;73(1-2):175–180. doi: 10.1016/0378-1097(92)90604-m. [DOI] [PubMed] [Google Scholar]
  2. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Dupont C., Clarke A. J. Dependence of lysozyme-catalysed solubilization of Proteus mirabilis peptidoglycan on the extent of O-acetylation. Eur J Biochem. 1991 Feb 14;195(3):763–769. doi: 10.1111/j.1432-1033.1991.tb15764.x. [DOI] [PubMed] [Google Scholar]
  6. Dupont C., Clarke A. J. Evidence for N----O acetyl migration as the mechanism for O acetylation of peptidoglycan in Proteus mirabilis. J Bacteriol. 1991 Jul;173(14):4318–4324. doi: 10.1128/jb.173.14.4318-4324.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eisenbeis S. J., Parker J. Strains of Escherichia coli carrying the structural gene for histidyl-tRNA synthetase on a high copy-number plasmid. Mol Gen Genet. 1981;183(1):115–122. doi: 10.1007/BF00270148. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Fleischmann R. D., Adams M. D., White O., Clayton R. A., Kirkness E. F., Kerlavage A. R., Bult C. J., Tomb J. F., Dougherty B. A., Merrick J. M. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995 Jul 28;269(5223):496–512. doi: 10.1126/science.7542800. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hawkey P. M. Providencia stuartii: a review of a multiply antibiotic-resistant bacterium. J Antimicrob Chemother. 1984 Mar;13(3):209–226. doi: 10.1093/jac/13.3.209. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Kaniga K., Delor I., Cornelis G. R. A wide-host-range suicide vector for improving reverse genetics in gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica. Gene. 1991 Dec 20;109(1):137–141. doi: 10.1016/0378-1119(91)90599-7. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. McHale P. J., Walker F., Scully B., English L., Keane C. T. Providencia stuartii infections: a review of 117 cases over an eight year period. J Hosp Infect. 1981 Jun;2(2):155–165. doi: 10.1016/0195-6701(81)90024-4. [DOI] [PubMed] [Google Scholar]
  17. Miller V. L., Mekalanos J. J. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. 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]
  22. Warren J. W. Providencia stuartii: a common cause of antibiotic-resistant bacteriuria in patients with long-term indwelling catheters. Rev Infect Dis. 1986 Jan-Feb;8(1):61–67. doi: 10.1093/clinids/8.1.61. [DOI] [PubMed] [Google Scholar]
  23. 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]

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