1887

Microbiology Society logo

Volume 143, Issue 10

Transcriptional control of several aerobically induced cytochrome structural genes in Free

Abstract

To decipher how the synthesis of energy-transducing enzymes responds to environmental cues, the response of three aerobic cytochrome gene promoters was analysed under different conditions. Two of these promoters are upstream of structural genes ( and ) for individual subunits of the cytochrome respiratory complex. The third promoter is that for the operon, which encodes two -type cytochromes of unknown function, cytochrome and CycG. Primer extension analysis identified a single oxygen-responsive transcription start site for each gene. Utilizing operon fusions to as a measure of promoter activity, transcription from the and promoters was approximately twofold higher when cells were grown at high (30%) oxygen tensions than under low (2%) oxygen or anaerobic (photosynthetic) conditions. Analysis of promoter function using specific host mutations indicated that loss of the FNR homologue, FnrL, causes a small, but reproducible, increase in and transcription when cells are grown at 2% oxygen. However, neither the ΔFnrL mutation nor alterations in sequences related to a consensus target site for the FNR protein increased function of any of these three promoters to that seen under aerobic conditions in wild-type cells. From this we conclude that FnrL is not solely responsible for reduced transcription of these three aerobic cytochrome genes under low oxygen or anaerobic conditions. When activity of these three promoters was monitored after cells were shifted from anaerobic (photosynthetic) conditions to a 30% oxygen atmosphere, it took several cell doublings for LacZ levels to increase to those found in steady-state 30% oxygen cultures. From these results, it appears that activity of these promoters is also regulated by a stable molecule whose synthesis or function responds slowly to the presence of high oxygen tensions.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): anaerobic regulation , cytochromes , gene expression , oxygen regulation and transcriptional control
© Society for General Microbiology 1997
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-10-3101
1997-10-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/10/mic-143-10-3101.html?itemId=/content/journal/micro/10.1099/00221287-143-10-3101&mimeType=html&fmt=ahah

References

  1. Bagdasarian M., Timmis K. N. 1982; Host: Vector systems for gene cloning in pseudomonas . Curr Top Microbiol Immunol 96:47–67
     [Google Scholar]
  2. Cao J., Shapleigh J., Gennis R., Revzin A., Ferguson-Miller S. 1991; The gene encoding cytochrome c oxidase subunit ii from rhodobacter sphaeroides; comparison of the deduced amino acid sequences of corresponding peptides from other species. Gene 101:133–137
     [Google Scholar]
  3. Cao J., Hosier J., Shapleigh J., Revzin A., Ferguson-Miller S. 1992; Cytochrome aa3 of rhodobacter sphaeroides as a model for mitochondrial cytochrome c oxidase: the coxII/coxIII operon codes for structural and assembly proteins homologous to those in yeast. J Biol Chem 267:24273–24278
     [Google Scholar]
  4. Chory J., Donohue T. J., Varga A. R., Staehlin L. A., Kaplan S. 1984; Induction of photosynthetic membranes of rhodopseudomonas sphaeroides: Biochemical and morphological studies. J Bacteriol 159:540–554
     [Google Scholar]
  5. Cohen-Bazire G., Sistrom W. R., Stanier R. Y. 1957; Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cell Comp Physiol 49:25–68
     [Google Scholar]
  6. Cuypers H., Zumft W. G. 1993; Anaerobic control of denitrification in pseudomonas stutzeri escapes mutagenesis of an fnr-like gene. J Bacteriol 175:7236–7246
     [Google Scholar]
  7. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the vax. Nucleic Acids Res 12:387–395
     [Google Scholar]
  8. Donohue T. J., Kaplan S. 1991; Genetic techniques in the rhodospirillaceae. Methods Enzymol 204:459–485
     [Google Scholar]
  9. Donohue T. J., McEwan A. G., Kaplan S. 1986; Cloning, dna sequence, and expression of the rhodobacter sphaeroides cytochrome c 2 gene. J 168:962–972
     [Google Scholar]
  10. Donohue T. J., McEwan A. G., Van Doren S., Crofts A., Kaplan S. 1988; Phenotypic and genetic characterization of cytochrome c 2 deficient mutants of rhodobacter sphaeroides . Biochemistry 27:1918–1925
     [Google Scholar]
  11. Eraso J. M., Kaplan S. 1994; prrA, a putative response regulatory protein involved in oxygen regulation of photosynthesis gene expression in Rhodobacter sphaeroides.. J Bacteriol 176:32–43
     [Google Scholar]
  12. Flory J. E., Donohue T. J. 1995; Organization and expression of the rhodobacter sphaeroides cycFG operon. J Bacteriol 177:4311–4320
     [Google Scholar]
  13. Garcfa-Horsman J. A., Barquera B., Rumbley J., Ma j., Gennis R. B. The superfamily of heme-copper respiratory oxidases.. J Bacteriol 176:5587–5600
     [Google Scholar]
  14. Garcfa-Horsman J. A., Berry E., Shapleigh J. P., Alben J. O., Gennis R. B. 1994; A novel cytochrome c oxidase from rhodobacter sphaeroides that lacks cuA . Biochemistry 33:3113–3119
     [Google Scholar]
  15. Gomelsky M., Kaplan S. 1995; Genetic evidence that PpsR from rhodobacter sphaeroides 2.4.1 functions as a repressor of puc and bchf expression. J Bacteriol 177:1634–1637
     [Google Scholar]
  16. Green J., Guest J. R. 1994; Regulation of transcription at the ndh promoter of escherichia coli by fnr and novel factors. Mol Microbiol 12:433–444
     [Google Scholar]
  17. Luchi S., Chepuri V., Fu H. A., Gennis R. B., Lin E. C. C. 1990; Requirement for terminal cytochromes in generation of the signal for the arc regulatory system in Escherichia coli: Study using deletions and lac fusions to cyo and cyd . J Bacteriol 172:6020–6025
     [Google Scholar]
  18. Karls R., Schulz V., Jovanovich S. B., Flynn S., Pak A., Reznikoff W. S. 1989; Pseudorevertants of a lac promoter mutation reveal overlapping nascent promoters. Nucleic Acids Res 17:3927–3949
     [Google Scholar]
  19. Kunkel T. A. 1985; Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
     [Google Scholar]
  20. Lazazzera B. A., Bates D. M., Kiley P. J. 1993; The activity of the Escherichia coli transcription factor fnr is regulated by a change in oligomeric state. Genes Dev 7:1993–2005
     [Google Scholar]
  21. Lee J. K., Kaplan S. 1992; cis-acting regulatory elements involved in oxygen and light control of puc operon transcription in Rhodobacter sphaeroides . J Bacteriol 174:1146–1157
     [Google Scholar]
  22. Markwell M. A. K., Haas S. M., Bieber L. L., Tolbert N. E. 1978; A modification of the lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210
     [Google Scholar]
  23. Minagawa J., Nakamura H., Yamato I., Anraku Y. 1990; Transcriptional regulation of the cytochrome b 562—o complex in Escherichia Coli . J Biol Chem 265:11198–11203
     [Google Scholar]
  24. Moshiri F., Smith E. G., Taormino J. P., Maier R. J. 1991; Transcriptional regulation of cytochrome d in nitrogen-fixing Azotobacter Vinelandii . J Biol Chem 266:23169–23174
     [Google Scholar]
  25. Penfold R. J., Pemberton J. M. 1994; Sequencing, chromosomal inactivation, and functional expression in Escherichia Coli of ppsR, a gene which represses carotenoid and bacteriochlorophyll synthesis in Rhodobacter Sphaeroides . J Bacteriol 176:2869–2876
     [Google Scholar]
  26. Prentki P., Krisch H. M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313
     [Google Scholar]
  27. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  28. Schilke B. A., Donohue T. J. 1992; δ-Aminolevulinate couples cycA transcription to changes in heme availability in Rhodobacter sphaeroides.. J Mol Biol 226:101–115
     [Google Scholar]
  29. Shapleigh J. P., Gennis R. B. 1992; Cloning, sequencing and deletion from the chromosome of the gene encoding subunit i of the aa 3-type cytochrome c oxidase of Rhodobacter Spbaeroides . Mol Microbiol 6:635–642
     [Google Scholar]
  30. Sharrocks A. D., Green J., Guest J. R. 1991; FNR activates and represses transcription In Vitro.. Proc R Soc Lond B 245:219–226
     [Google Scholar]
  31. Simon R., Priefer U., Puhler A. 1983; A broad host range mobilization system for in vivo genetic engineering: Transposon mutagenesis in gram-negative bacteria. Bio/Technology 1:784–791
     [Google Scholar]
  32. Spiro S., Guest J. R. 1991; Adaptive responses to oxygen limitation in Escherichia Coli.. Trends Biochem Sci 16:310–314
     [Google Scholar]
  33. Spiro S., Gaston K. L., Bell A. I., Roberts R. E., Busby S. J. W., Guest J. R. 1990; Interconversion of the DNA-binding specificities of two related transcription regulators, crp and fnr. Mol Microbiol 4:1831–1838
     [Google Scholar]
  34. Tosques I. E., Shi J., Shapleigh J. R. 1996; Cloning and characterization of nnrR, whose product is required for the expression of proteins involved in nitric oxide metabolism in Rhodobacter Spbaeroides 2.4.3. J Bacteriol 178:4958–4964
     [Google Scholar]
  35. Trumpower B. L., Gennis R. B. 1994; Energy transduction by cytochrome complexes in mitochondrial and bacterial respiration : The enzymology of coupling electron transfer reactions to transmembrane proton translocation. Annu Rev Biochem 63:675–716
     [Google Scholar]
  36. Zeilstra-Ryalls J. H., Kaplan S. 1995; Aerobic and anaerobic regulation in Rhodobacter Spbaeroides 2.4.1: The role of the fnrL gene. J Bacteriol 177:6422–6431
     [Google Scholar]
  37. Zhu Y. S., Kaplan S. 1985; Effects of light, oxygen, and substrates on steady-state levels of mrna coding for ribulose- 1,5-bisphosphate carboxylase and light-harvesting and reaction center polypeptides in Rhodopseudomonas Spbaeroides . J Bacteriol 162:925–932
     [Google Scholar]
  38. Ziegelhoffer E., Kiley P. J. 1995; In vitro analysis of a constitutively active mutant form of the Escherichia coli global transcription factor FNR. J Mol Biol 245:351–361
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-10-3101
Loading
Transcriptional control of several aerobically induced cytochrome structural genes in Rhodobacter sphaeroides
Microbiology 143, 3101 (1997); https://doi.org/10.1099/00221287-143-10-3101
/content/journal/micro/10.1099/00221287-143-10-3101
/content/journal/micro/10.1099/00221287-143-10-3101
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error