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. 1995 Mar 11;23(5):819–826. doi: 10.1093/nar/23.5.819

Selectivity of the Escherichia coli RNA polymerase E sigma 38 for overlapping promoters and ability to support CRP activation.

A Kolb 1, D Kotlarz 1, S Kusano 1, A Ishihama 1
PMCID: PMC306765  PMID: 7708498

Abstract

A series of gal promoter mutants has been used to compare the in vitro selectivities of the two forms of Escherichia coli RNA polymerase, E sigma 38 and E sigma 70. In the absence of the CRP-cAMP complex, E sigma 38 shows a strong preference for the ga/P1 promoter, whereas E sigma 70 preferentially initiates transcription from the ga/P2 promoter. E sigma 38 selectivity is not affected by the nature and position of the upstream sequences or by the phasing between synthetic upstream curved sequences and the -10 regions. In fact, all effects of mutations in the extended -10 region can be accounted for without evoking strong new sequence preferences for E sigma 38. Finally, both E sigma 38 and E sigma 70 initiate transcription from the ga/P1 promoter in the presence of CRP-cAMP complex and support direct cAMP-CRP activation at several CRP-dependent promoters.

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Selected References

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  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Attey A., Belyaeva T., Savery N., Hoggett J., Fujita N., Ishihama A., Busby S. Interactions between the cyclic AMP receptor protein and the alpha subunit of RNA polymerase at the Escherichia coli galactose operon P1 promoter. Nucleic Acids Res. 1994 Oct 25;22(21):4375–4380. doi: 10.1093/nar/22.21.4375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bingham A. H., Ponnambalam S., Chan B., Busby S. Mutations that reduce expression from the P2 promoter of the Escherichia coli galactose operon. Gene. 1986;41(1):67–74. doi: 10.1016/0378-1119(86)90268-4. [DOI] [PubMed] [Google Scholar]
  4. Bracco L., Kotlarz D., Kolb A., Diekmann S., Buc H. Synthetic curved DNA sequences can act as transcriptional activators in Escherichia coli. EMBO J. 1989 Dec 20;8(13):4289–4296. doi: 10.1002/j.1460-2075.1989.tb08615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Busby S., Aiba H., de Crombrugghe B. Mutations in the Escherichia coli operon that define two promoters and the binding site of the cyclic AMP receptor protein. J Mol Biol. 1982 Jan 15;154(2):211–227. doi: 10.1016/0022-2836(82)90061-4. [DOI] [PubMed] [Google Scholar]
  6. Busby S., Kotlarz D., Buc H. Deletion mutagenesis of the Escherichia coli galactose operon promoter region. J Mol Biol. 1983 Jun 25;167(2):259–274. doi: 10.1016/s0022-2836(83)80335-0. [DOI] [PubMed] [Google Scholar]
  7. Busby S., Truelle N., Spassky A., Dreyfus M., Buc H. The selection and characterisation of two novel mutations in the overlapping promoters of the Escherichia coli galactose operon. Gene. 1984 May;28(2):201–209. doi: 10.1016/0378-1119(84)90257-9. [DOI] [PubMed] [Google Scholar]
  8. Chan B., Spassky A., Busby S. The organization of open complexes between Escherichia coli RNA polymerase and DNA fragments carrying promoters either with or without consensus -35 region sequences. Biochem J. 1990 Aug 15;270(1):141–148. doi: 10.1042/bj2700141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Espinosa-Urgel M., Tormo A. Sigma s-dependent promoters in Escherichia coli are located in DNA regions with intrinsic curvature. Nucleic Acids Res. 1993 Aug 11;21(16):3667–3670. doi: 10.1093/nar/21.16.3667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gaston K., Bell A., Kolb A., Buc H., Busby S. Stringent spacing requirements for transcription activation by CRP. Cell. 1990 Aug 24;62(4):733–743. doi: 10.1016/0092-8674(90)90118-x. [DOI] [PubMed] [Google Scholar]
  11. Gribskov M., Burgess R. R. Overexpression and purification of the sigma subunit of Escherichia coli RNA polymerase. Gene. 1983 Dec;26(2-3):109–118. doi: 10.1016/0378-1119(83)90180-4. [DOI] [PubMed] [Google Scholar]
  12. Hengge-Aronis R. Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E. coli. Cell. 1993 Jan 29;72(2):165–168. doi: 10.1016/0092-8674(93)90655-a. [DOI] [PubMed] [Google Scholar]
  13. Ishihama A. Protein-protein communication within the transcription apparatus. J Bacteriol. 1993 May;175(9):2483–2489. doi: 10.1128/jb.175.9.2483-2489.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Juang Y. L., Helmann J. D. A promoter melting region in the primary sigma factor of Bacillus subtilis. Identification of functionally important aromatic amino acids. J Mol Biol. 1994 Feb 4;235(5):1470–1488. doi: 10.1006/jmbi.1994.1102. [DOI] [PubMed] [Google Scholar]
  15. Kolb A., Buc H. Is DNA unwound by the cyclic AMP receptor protein? Nucleic Acids Res. 1982 Jan 22;10(2):473–485. doi: 10.1093/nar/10.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kolb A., Igarashi K., Ishihama A., Lavigne M., Buckle M., Buc H. E. coli RNA polymerase, deleted in the C-terminal part of its alpha-subunit, interacts differently with the cAMP-CRP complex at the lacP1 and at the galP1 promoter. Nucleic Acids Res. 1993 Jan 25;21(2):319–326. doi: 10.1093/nar/21.2.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuhnke G., Krause A., Heibach C., Gieske U., Fritz H. J., Ehring R. The upstream operator of the Escherichia coli galactose operon is sufficient for repression of transcription initiated at the cyclic AMP-stimulated promoter. EMBO J. 1986 Jan;5(1):167–173. doi: 10.1002/j.1460-2075.1986.tb04192.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kumar A., Malloch R. A., Fujita N., Smillie D. A., Ishihama A., Hayward R. S. The minus 35-recognition region of Escherichia coli sigma 70 is inessential for initiation of transcription at an "extended minus 10" promoter. J Mol Biol. 1993 Jul 20;232(2):406–418. doi: 10.1006/jmbi.1993.1400. [DOI] [PubMed] [Google Scholar]
  19. Lange R., Hengge-Aronis R. The cellular concentration of the sigma S subunit of RNA polymerase in Escherichia coli is controlled at the levels of transcription, translation, and protein stability. Genes Dev. 1994 Jul 1;8(13):1600–1612. doi: 10.1101/gad.8.13.1600. [DOI] [PubMed] [Google Scholar]
  20. Lavigne M., Herbert M., Kolb A., Buc H. Upstream curved sequences influence the initiation of transcription at the Escherichia coli galactose operon. J Mol Biol. 1992 Mar 20;224(2):293–306. doi: 10.1016/0022-2836(92)90995-v. [DOI] [PubMed] [Google Scholar]
  21. Lederer H., Mortensen K., May R. P., Baer G., Crespi H. L., Dersch D., Heumann H. Spatial arrangement of sigma-factor and core enzyme of Escherichia coli RNA polymerase. A neutron solution scattering study. J Mol Biol. 1991 Jun 20;219(4):747–755. doi: 10.1016/0022-2836(91)90669-w. [DOI] [PubMed] [Google Scholar]
  22. Loewen P. C., Hengge-Aronis R. The role of the sigma factor sigma S (KatF) in bacterial global regulation. Annu Rev Microbiol. 1994;48:53–80. doi: 10.1146/annurev.mi.48.100194.000413. [DOI] [PubMed] [Google Scholar]
  23. Loewen P. C., von Ossowski I., Switala J., Mulvey M. R. KatF (sigma S) synthesis in Escherichia coli is subject to posttranscriptional regulation. J Bacteriol. 1993 Apr;175(7):2150–2153. doi: 10.1128/jb.175.7.2150-2153.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lonetto M., Gribskov M., Gross C. A. The sigma 70 family: sequence conservation and evolutionary relationships. J Bacteriol. 1992 Jun;174(12):3843–3849. doi: 10.1128/jb.174.12.3843-3849.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McCann M. P., Fraley C. D., Matin A. The putative sigma factor KatF is regulated posttranscriptionally during carbon starvation. J Bacteriol. 1993 Apr;175(7):2143–2149. doi: 10.1128/jb.175.7.2143-2149.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moyle H., Waldburger C., Susskind M. M. Hierarchies of base pair preferences in the P22 ant promoter. J Bacteriol. 1991 Mar;173(6):1944–1950. doi: 10.1128/jb.173.6.1944-1950.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mulvey M. R., Loewen P. C. Nucleotide sequence of katF of Escherichia coli suggests KatF protein is a novel sigma transcription factor. Nucleic Acids Res. 1989 Dec 11;17(23):9979–9991. doi: 10.1093/nar/17.23.9979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Newlands J. T., Gaal T., Mecsas J., Gourse R. L. Transcription of the Escherichia coli rrnB P1 promoter by the heat shock RNA polymerase (E sigma 32) in vitro. J Bacteriol. 1993 Feb;175(3):661–668. doi: 10.1128/jb.175.3.661-668.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nguyen L. H., Jensen D. B., Thompson N. E., Gentry D. R., Burgess R. R. In vitro functional characterization of overproduced Escherichia coli katF/rpoS gene product. Biochemistry. 1993 Oct 19;32(41):11112–11117. doi: 10.1021/bi00092a021. [DOI] [PubMed] [Google Scholar]
  30. Ponnambalam S., Spassky A., Busby S. Studies with the Escherichia coli galactose operon regulatory region carrying a point mutation that simultaneously inactivates the two overlapping promoters. Interactions with RNA polymerase and the cyclic AMP receptor protein. FEBS Lett. 1987 Jul 13;219(1):189–196. doi: 10.1016/0014-5793(87)81214-0. [DOI] [PubMed] [Google Scholar]
  31. Ponnambalam S., Webster C., Bingham A., Busby S. Transcription initiation at the Escherichia coli galactose operon promoters in the absence of the normal -35 region sequences. J Biol Chem. 1986 Dec 5;261(34):16043–16048. [PubMed] [Google Scholar]
  32. Richet E., Raibaud O. Supercoiling is essential for the formation and stability of the initiation complex at the divergent malEp and malKp promoters. J Mol Biol. 1991 Apr 5;218(3):529–542. doi: 10.1016/0022-2836(91)90699-7. [DOI] [PubMed] [Google Scholar]
  33. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  34. Tanaka K., Kusano S., Fujita N., Ishihama A., Takahashi H. Promoter determinants for Escherichia coli RNA polymerase holoenzyme containing sigma 38 (the rpoS gene product). Nucleic Acids Res. 1995 Mar 11;23(5):827–834. doi: 10.1093/nar/23.5.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tanaka K., Takayanagi Y., Fujita N., Ishihama A., Takahashi H. Heterogeneity of the principal sigma factor in Escherichia coli: the rpoS gene product, sigma 38, is a second principal sigma factor of RNA polymerase in stationary-phase Escherichia coli. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3511–3515. doi: 10.1073/pnas.90.8.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]

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