Abstract
The regulatory elements involved in expression of the gene (fdhF) for the selenopolypeptide of formate dehydrogenase and of a gene (or transcriptional unit) (hyd) specifically responsible for the formation of the gas-evolving hydrogenase (hydrogenase 3) in Escherichia coli were investigated. Formate (or a product of it) is required for expression of both systems since in a pyruvate-formate-lyase deficient mutant induction occurs only when formate is supplemented externally. Under this condition, formate can partially overcome repression by nitrate. The transcription of both the fdhF gene and the hydrogenase-3-encoding systems is independent of the presence of a wild-type fnr gene when formate is present, supporting the view that the Fnr effect on the formation of the formate-hydrogen-lyase pathway is indirect. Mutations blocking the synthesis of a functional molybdenum cofactor also had no major affect on fdhF and hyd expression.
The nucleotide sequence of the 5′ flanking region of the fdhF gene was determined and the transcription start point of the fdhF gene was localized by nuclease S1 mapping. Nuclease Bal31 generated deletion clones were constructed and the regulation of their expression was studied. Anaerobic expression and induction by formate depended on the presence of a stretch of approximately 185 nucleotides upstream of the translation start. Elements mediating formate induction and oxygen or nitrate repression could not be separated physically. The regulatory features of the fdhF upstream region bear striking resemblance to systems whose expression are dependent upon upstream activating elements.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aiba H, Adya S, de Crombrugghe B (1981) Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
Alvarez-Morales A, Betancourt-Alvarez M, Kaluza K, Hennecke H (1986) Activation of the Bradyrhizobium japonicum nifH and nifDK operons is dependent on promoter-upstream DNA sequences. Nucleic Acids Res 14:4007–4027
Ballantine SP, Boxer DH (1985) Nickel-containing hydrogenase isoenzymes from anaerobically grown Escherichia coli K-12. J. Bacteriol 163:454–459
Ballantine SP, Boxer DH (1986) Isolation and characterization of a soluble active fragment of hydrogenase isoenzyme 2 from the membranes of anaerobically grown Escherichia coli. Eur J Biochem 156:277–284
Begg IA, Whyte JN, Haddock BA (1977) The identification of mutants of Escherichia coli deficient in formate dehydrogenase and nitrate reductase activities using dye indicator plates. FEMS Microbiol Lett 2:47–50
Bonnefoy V, Pascal MC, Ratouchniak J, Chippaux M (1986) Autoregulation of the nar operon encoding nitrate reductase in Escherichia coli. Mol Gen Genet 204:180–184
Buck M, Miller S, Drummond M, Dixon R (1986) Upstream activator sequences are present in the promoters of nitrogen fixation genes. Nature (Lond) 320:374–378
Casadaban MJ, Cohen SN (1979) Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci USA 76:4430–4533
Casadaban MJ, Chou J, Cohen SN (1980) In vitro gene fusions that join an enzymatically active β-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol 143:971–980
Chippaux M, Gindici D, Abou-Jaoude A, Casse F, Pascal MC (1978) A mutation leading to the total lack of nitrite reductase activity in Escherichia coli K-12. Mol Gen Genet 160:225–229
Chippaux M, Bonnefoy-Orth V, Ratouchniak J, Pascal MC (1981) Operon fusions in the nitrate reductase operon and study of the control gene nirR in Escherichia coli. Mol Gen Genet 182:477–479
Cole JA, Wimpenny JWT (1966) The interrelationships of low redox potential cytochrome C552 and hydrogenase in facultative anaerobes. Biochim Biophys Acta 128:419–425
Csonka LN, Clark AJ (1980) Construction of an Hfr strain useful for transferring recA mutations between Escherichia coli strains. J Bacteriol 143:529–530
Gray CT, Gest H (1965) Biological formation of molecular hydrogen. Science 148:186–192
Ihara M, Oda Y, Yamamoto K (1985) Convenient construction of strains useful for transducing recA mutations with bacteriophage P1. FEMS Microbiol Lett 30:33–35
Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Improved technique utilising nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1:3–8
Kuritzkes DR, Zhang XY, Lin ECC (1984) Use of ϕ (glp-lac) in studies of respiratory regulation of the Escherichia coli anaerobic sn-glycerol-3-phosphate dehydrogenase genes (glp AB). J Bacteriol 157:591–598
Lambden PR, Guest JR (1976) Mutants of Escherichia coli K-12 unable to use fumarate as an anaerobic electron acceptor. J Gen Microbiol 97:145–160
Lee HJ, Patel P, Sankar P, Shanmugam KT (1985) Isolation and characterization of mutant strains of Escherichia coli altered in H2 metabolism. J Bacteriol 162:344–352
Li S, Rabi T, De Moss J (1985) Delineation of two distinct regulatory domains in the 5′ region of the nar operon of Escherichia coli. J Bacteriol 164:25–32
MacGregor CH (1975) Synthesis of nitrate reductase components in chlorate resistant mutants of Escherichia coli. J Bacteriol 121:1117–1121
MacGregor CH, Schnaitman CA (1971) Alteration in the cytoplasmic membrane proteins of various chlorate resistant mutants of Escherichia coli. J Bacteriol 108:564–570
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavage. Methods Enzymol 65:499–560
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Mizuno T, Mizushima S (1986) Characterization by deletion and localized mutagenesis in vitro of the promoter region of the Escherichia coli ompC gene and importance of the upstream DNA domain in positive regulation by the ompR protein. J Bacteriol 168:86–95
Newman M, Cole JA (1978) The chromosomal location and pleiotropic effects of mutations of the nirA + gene of Escherichia coli K-12: The essential role of nirA + in nitrite reduction and in other anaerobic redox reactions. J Gen Microbiol 106:1–12
Pascal MC, Burini JF, Ratouchniak J, Chippaux M (1982) Regulation of the nitrate reductase operon: Effect of mutations in chlA, B, D, and E genes. Mol Gen Genet 188:103–106
Pecher A, Zinoni F, Jatisatienr C, Wirth R, Hennecke H, Böck A (1983) On the redox control of synthesis of anaerobically induced enzymes in enterobacteriaceae. Arch Microbiol 136:131–136
Pecher A, Zinoni F, Böck A (1985) The seleno-polypeptide of formic dehydrogenase (formate-hydrogen-lyase linked) from Escherichia coli: genetic analysis. Arch Microbiol 141:359–363
Peck HD, Gest H (1957) Formic dehydrogenase and the hydrogenlyase enzyme complex in the coli-aerogenes group. J Bacteriol 73:706–721
Reitzer LJ, Magasanik B (1986) Transcription of glnA in E. coli is stimulated by activator bound to sites far from the promoter. Cell 45:785–792
Sawers RG, Boxer DH (1986) Purification and properties of membrane-bound hydrogenase isoenzyme 1 from anaerobically grown Escherichia coli K12. Eur J Biochem 156:265–275
Sawers RG, Ballantine SP, Boxer DH (1985) Differential expression of hydrogenase isoenzymes in Escherichia coli K-12: Evidence for a third isoenzyme. J Bacteriol 164:1324–1331
Sawers RG, Jamieson DJ, Higgins CF, Boxer DH (1986) Characterization and physiological roles of membrane-bound hydrogenase isoenzymes from Salmonella typhimurium. J Bacteriol 168:398–404
Smith MW, Neidhardt FC (1983) Proteins induced by anaerobiosis in Escherichia coli. J Bacteriol 154:336–343
Sollner-Webb B, Reeder RH (1979) The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell 18:485–499
Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stewart V, MacGregor C (1982) Nitrate reductase in Escherichia coli K-12: Involvement of chlC, chlE and chlG loci. J Bacteriol 151:788–799
Varenne S, Casse F, Chippaux, Pascal MC (1975) A mutant of Escherichia coli deficient in pyruvate formate lyase. Mol Gen Genet 141:181–184
Viera J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with universal primers. Gene 19:259–268
Waugh R, Boxer DH (1986) Pleiotropic hydrogenase mutants of Escherichia coli K-12: growth in the presence of nickel can restore hydrogenase activity. Biochimie 68:157–166
Weaver FR, Weissmann CH (1979) Mapping of RNA by a modification of the Berk-Sharp procedure: the 5′ termini of 15S β-globin mRNA precursor and mature 10S β-globin mRNA have identical map coordinates. Nucleic Acids Res 7:1175–1193
Wich G, Hummel H, Jarsch M, Bär U, Böck A (1986) Transcription signals for stable RNA genes in Methanococcus. Nucleic Acids Res 14:2459–2479
Wu LF, Mandrand-Berthelot MA (1986) Genetic and physiological characterization of new Escherichia coli mutants impaired in hydrogenase activity. Biochimie 68:167–179
Zinoni F, Beier A, Pecher A, Wirth R, Böck A (1984) Regulation of the synthesis of hydrogenase (formate-hydrogen-lyase linked) of E. coli. Arch Microbiol 139:299–304
Zinoni F, Birkmann A, Stadtman TC, Böck A (1986) Nucleotide sequence and expression of the selenocysteine containing polypeptide of formate dehydrogenase (formate-hydrogenlyase-linked) from Escherichia coli. Proc Natl Acad Sci USA 83:4650–4654
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Birkmann, A., Zinoni, F., Sawers, G. et al. Factors affecting transcriptional regulation of the formate-hydrogen-lyase pathway of Escherichia coli . Arch. Microbiol. 148, 44–51 (1987). https://doi.org/10.1007/BF00429646
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00429646