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. 1991 Dec;173(23):7501–7510. doi: 10.1128/jb.173.23.7501-7510.1991

Suppressor mutations in rpoA suggest that OmpR controls transcription by direct interaction with the alpha subunit of RNA polymerase.

J M Slauch 1, F D Russo 1, T J Silhavy 1
PMCID: PMC212516  PMID: 1657891

Abstract

We have isolated mutations in rpoA, the gene encoding the alpha subunit of RNA polymerase, that specifically affect transcriptional control by OmpR and EnvZ, the two-component regulatory system that controls porin gene expression in Escherichia coli. Characterization of these mutations and a previously isolated rpoA allele suggests that both positive and negative regulation of porin gene transcription involves a direct interaction between OmpR and RNA polymerase through the alpha subunit. Several of the rpoA mutations cluster in the carboxy-terminal portion of the alpha protein, further suggesting that it is this domain of alpha that is involved in interaction with OmpR and perhaps other transcriptional regulators as well.

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Selected References

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  1. Adhya S., Garges S. Positive control. J Biol Chem. 1990 Jul 5;265(19):10797–10800. [PubMed] [Google Scholar]
  2. Aiba H., Nakasai F., Mizushima S., Mizuno T. Evidence for the physiological importance of the phosphotransfer between the two regulatory components, EnvZ and OmpR, in osmoregulation in Escherichia coli. J Biol Chem. 1989 Aug 25;264(24):14090–14094. [PubMed] [Google Scholar]
  3. Bedwell D. M., Nomura M. Feedback regulation of RNA polymerase subunit synthesis after the conditional overproduction of RNA polymerase in Escherichia coli. Mol Gen Genet. 1986 Jul;204(1):17–23. doi: 10.1007/BF00330181. [DOI] [PubMed] [Google Scholar]
  4. Berman M. L., Jackson D. E. Selection of lac gene fusions in vivo: ompR-lacZ fusions that define a functional domain of the ompR gene product. J Bacteriol. 1984 Aug;159(2):750–756. doi: 10.1128/jb.159.2.750-756.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brickman E., Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and phi80 transducing phages. J Mol Biol. 1975 Aug 5;96(2):307–316. doi: 10.1016/0022-2836(75)90350-2. [DOI] [PubMed] [Google Scholar]
  6. Bushman F. D., Ptashne M. Turning lambda Cro into a transcriptional activator. Cell. 1988 Jul 15;54(2):191–197. doi: 10.1016/0092-8674(88)90551-x. [DOI] [PubMed] [Google Scholar]
  7. Bushman F. D., Shang C., Ptashne M. A single glutamic acid residue plays a key role in the transcriptional activation function of lambda repressor. Cell. 1989 Sep 22;58(6):1163–1171. doi: 10.1016/0092-8674(89)90514-x. [DOI] [PubMed] [Google Scholar]
  8. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  9. Case C. C., Bukau B., Granett S., Villarejo M. R., Boos W. Contrasting mechanisms of envZ control of mal and pho regulon genes in Escherichia coli. J Bacteriol. 1986 Jun;166(3):706–712. doi: 10.1128/jb.166.3.706-712.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Christie G. E., Haggård-Ljungquist E., Feiwell R., Calendar R. Regulation of bacteriophage P2 late-gene expression: the ogr gene. Proc Natl Acad Sci U S A. 1986 May;83(10):3238–3242. doi: 10.1073/pnas.83.10.3238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Forst S., Delgado J., Rampersaud A., Inouye M. In vivo phosphorylation of OmpR, the transcription activator of the ompF and ompC genes in Escherichia coli. J Bacteriol. 1990 Jun;172(6):3473–3477. doi: 10.1128/jb.172.6.3473-3477.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garrett S., Silhavy T. J. Isolation of mutations in the alpha operon of Escherichia coli that suppress the transcriptional defect conferred by a mutation in the porin regulatory gene envZ. J Bacteriol. 1987 Apr;169(4):1379–1385. doi: 10.1128/jb.169.4.1379-1385.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Giffard P. M., Booth I. R. The rpoA341 allele of Escherichia coli specifically impairs the transcription of a group of positively-regulated operons. Mol Gen Genet. 1988 Sep;214(1):148–152. doi: 10.1007/BF00340193. [DOI] [PubMed] [Google Scholar]
  14. Hall M. N., Silhavy T. J. The ompB locus and the regulation of the major outer membrane porin proteins of Escherichia coli K12. J Mol Biol. 1981 Feb 15;146(1):23–43. doi: 10.1016/0022-2836(81)90364-8. [DOI] [PubMed] [Google Scholar]
  15. Hall M. N., Silhavy T. J. Transcriptional regulation of Escherichia coli K-12 major outer membrane protein 1b. J Bacteriol. 1979 Nov;140(2):342–350. doi: 10.1128/jb.140.2.342-350.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hancock R. E., Reeves P. Bacteriophage resistance in Escherichia coli K-12: general pattern of resistance. J Bacteriol. 1975 Mar;121(3):983–993. doi: 10.1128/jb.121.3.983-993.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hawley D. K., McClure W. R. Mechanism of activation of transcription initiation from the lambda PRM promoter. J Mol Biol. 1982 May 25;157(3):493–525. doi: 10.1016/0022-2836(82)90473-9. [DOI] [PubMed] [Google Scholar]
  18. Hwang J. J., Gussin G. N. Interactions between Escherichia coli RNA polymerase and lambda repressor. Mutations in PRM affect repression of PR. J Mol Biol. 1988 Apr 20;200(4):735–739. doi: 10.1016/0022-2836(88)90484-6. [DOI] [PubMed] [Google Scholar]
  19. Igarashi K., Ishihama A. Bipartite functional map of the E. coli RNA polymerase alpha subunit: involvement of the C-terminal region in transcription activation by cAMP-CRP. Cell. 1991 Jun 14;65(6):1015–1022. doi: 10.1016/0092-8674(91)90553-b. [DOI] [PubMed] [Google Scholar]
  20. Igo M. M., Slauch J. M., Silhavy T. J. Signal transduction in bacteria: kinases that control gene expression. New Biol. 1990 Jan;2(1):5–9. [PubMed] [Google Scholar]
  21. Kretz K. A., Carson G. S., O'Brien J. S. Direct sequencing from low-melt agarose with Sequenase. Nucleic Acids Res. 1989 Jul 25;17(14):5864–5864. doi: 10.1093/nar/17.14.5864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lombardo M. J., Bagga D., Miller C. G. Mutations in rpoA affect expression of anaerobically regulated genes in Salmonella typhimurium. J Bacteriol. 1991 Dec;173(23):7511–7518. doi: 10.1128/jb.173.23.7511-7518.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Malan T. P., Kolb A., Buc H., McClure W. R. Mechanism of CRP-cAMP activation of lac operon transcription initiation activation of the P1 promoter. J Mol Biol. 1984 Dec 25;180(4):881–909. doi: 10.1016/0022-2836(84)90262-6. [DOI] [PubMed] [Google Scholar]
  24. Martin K., Huo L., Schleif R. F. The DNA loop model for ara repression: AraC protein occupies the proposed loop sites in vivo and repression-negative mutations lie in these same sites. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3654–3658. doi: 10.1073/pnas.83.11.3654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Matsuyama S., Mizuno T., Mizushima S. Interaction between two regulatory proteins in osmoregulatory expression of ompF and ompC genes in Escherichia coli: a novel ompR mutation suppresses pleiotropic defects caused by an envZ mutation. J Bacteriol. 1986 Dec;168(3):1309–1314. doi: 10.1128/jb.168.3.1309-1314.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Matsuyama S., Mizushima S. Novel rpoA mutation that interferes with the function of OmpR and EnvZ, positive regulators of the ompF and ompC genes that code for outer-membrane proteins in Escherichia coli K12. J Mol Biol. 1987 Jun 20;195(4):847–853. doi: 10.1016/0022-2836(87)90489-x. [DOI] [PubMed] [Google Scholar]
  27. McCarthy J. E., Gualerzi C. Translational control of prokaryotic gene expression. Trends Genet. 1990 Mar;6(3):78–85. doi: 10.1016/0168-9525(90)90098-q. [DOI] [PubMed] [Google Scholar]
  28. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ostrow K. S., Silhavy T. J., Garrett S. cis-acting sites required for osmoregulation of ompF expression in Escherichia coli K-12. J Bacteriol. 1986 Dec;168(3):1165–1171. doi: 10.1128/jb.168.3.1165-1171.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rowland G. C., Giffard P. M., Booth I. R. phs Locus of Escherichia coli, a mutation causing pleiotropic lesions in metabolism, is an rpoA allele. J Bacteriol. 1985 Nov;164(2):972–975. doi: 10.1128/jb.164.2.972-975.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schnaitman C. A., McDonald G. A. Regulation of outer membrane protein synthesis in Escherichia coli K-12: deletion of ompC affects expression of the OmpF protein. J Bacteriol. 1984 Aug;159(2):555–563. doi: 10.1128/jb.159.2.555-563.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shih M. C., Gussin G. N. Differential effects of mutations on discrete steps in transcription initiation at the lambda PRE promoter. Cell. 1983 Oct;34(3):941–949. doi: 10.1016/0092-8674(83)90551-2. [DOI] [PubMed] [Google Scholar]
  33. Slauch J. M., Garrett S., Jackson D. E., Silhavy T. J. EnvZ functions through OmpR to control porin gene expression in Escherichia coli K-12. J Bacteriol. 1988 Jan;170(1):439–441. doi: 10.1128/jb.170.1.439-441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Slauch J. M., Silhavy T. J. Genetic analysis of the switch that controls porin gene expression in Escherichia coli K-12. J Mol Biol. 1989 Nov 20;210(2):281–292. doi: 10.1016/0022-2836(89)90330-6. [DOI] [PubMed] [Google Scholar]
  35. Slauch J. M., Silhavy T. J. cis-acting ompF mutations that result in OmpR-dependent constitutive expression. J Bacteriol. 1991 Jul;173(13):4039–4048. doi: 10.1128/jb.173.13.4039-4048.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Straney S. B., Crothers D. M. Lac repressor is a transient gene-activating protein. Cell. 1987 Dec 4;51(5):699–707. doi: 10.1016/0092-8674(87)90093-6. [DOI] [PubMed] [Google Scholar]
  37. Su W., Porter S., Kustu S., Echols H. DNA-looping and enhancer activity: association between DNA-bound NtrC activator and RNA polymerase at the bacterial glnA promoter. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5504–5508. doi: 10.1073/pnas.87.14.5504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sunshine M. G., Sauer B. A bacterial mutation blocking P2 phage late gene expression. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2770–2774. doi: 10.1073/pnas.72.7.2770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tan S., Richmond T. J. DNA binding-induced conformational change of the yeast transcriptional activator PRTF. Cell. 1990 Jul 27;62(2):367–377. doi: 10.1016/0092-8674(90)90373-m. [DOI] [PubMed] [Google Scholar]
  40. Taylor R. K., Garrett S., Sodergren E., Silhavy T. J. Mutations that define the promoter of ompF, a gene specifying a major outer membrane porin protein. J Bacteriol. 1985 Jun;162(3):1054–1060. doi: 10.1128/jb.162.3.1054-1060.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wandersman C., Moreno F., Schwartz M. Pleiotropic mutations rendering Escherichia coli K-12 resistant to bacteriophage TP1. J Bacteriol. 1980 Sep;143(3):1374–1383. doi: 10.1128/jb.143.3.1374-1383.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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