Summary
A strain of E. coli carrying a Mudl insertion leading to chlorate resistance was found to lack nitrate reductase and formate dehydrogenase activities, but to synthesize b-type cytochrome constitutively. Introduction of this insertion mutation into a strain bearing a fusion between the nitrate reductase operon (chl C, chl I) and the lac structural genes resulted in the constitutive expression of the lac genes of this last fusion. Identical results were found when the Mudl was eliminated promoting a deletion in the original insertion site. This mutation was located midway between gal and aro A, at the chl E locus. Study of a chl E strain already described revealed similar behaviour. Absence of nitrate reductase activity in these strains which constitutively express the structural genes of the nitrate reductase operon was tentatively attributed to the simultaneous lack of a cofactor of the nitrate reductase terminal enzyme, possibly cofactor Mo-X, and of a repressor of the operon.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K 12, Edition 6. Microbiol Rev 44:1–56
Bonnefoy-Orth V, Lepelletier M, Pascal MC, Chippaux M (1981) Nitrate reductase and cytochrome bNitrate Reductase structural genes as parts of the nitrate reductase operon. Mol Gen Genet 181:535–540
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 sequence. Proc Natl Acad Sci USA 76:4530–4533
Chippaux M, Bonnefoy-Orth V, Ratouchniak J, Pascal MC (1981) Operon fusion in the nitrate reductase operon and study of the control gene nir R in Escherichia coli. Mol Gen Genet 182:477–479
Chippaux M, Forget P (1972) Mesure de l'activité hydrogénase. Biochimie 54:1217–1219
Chippaux M, Giudici D, Abou-Jaoudé 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
Glaser JH, De Moss JA (1971) Phenotypic restoration by molybdate of nitrate reductase activity in chl D mutants of Escherichia coli. J Bacteriol 108:854–860
Guymon ED, Spence JT (1966) The reduction of nitrate by molybdenum (V). Phys Chem 70:1964–1969
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin reagent. J Biol Chem 193:265–275
Mac Gregor CH (1975) Synthesis of nitrate reductase components in chlorate resistant mutants of Escherichia coli. J Bacteriol 121:1117–1121
Mac Gregor CH, Schnaitman CA (1971) Alterations in the cytoplasmic membrane proteins of various chlorate resistant mutants of Escherichia coli. J Bacteriol 108:564–570
Mac Gregor CH, Schnaitman CA (1973) Reconstitution of nitrate reductase activity and formation of membrane particles from cytoplasmic extracts of chlorate resistant mutants of Escherichia coli. J Bacteriol 114:1164–1176
Miller J (1972) Experiments in molecular biology. Cold Spring Harbor Press, New York
Orth V, Chippaux M, Pascal MC (1980) A mutant defective in electron transfer to nitrate in Escherichia coli K 12. J Gen Microbiol 117:257–262
Pascal MC, Casse F, Chippaux M (1977) Localization of pfl gene by transductional study of the gal-aro A segment of the Salmonella typhimurium LT2 chromosome. Mol Gen Genet 150:331–334
Pichinoty F (1962) Action répressive de l'oxygène sur la biosynthèse de la fumarique reductase d'Aerobacter aerogenes. Experientia 18:257–263
Pichinoty F (1969) Recherche des activités formiate oxydase, hydrogène-lyase, hydrogénase et formiate déshydrogénase chez quelques Enterobacteriaceae. Ann Inst Pasteur 117:3–15
Pichinoty F, Chippaux M (1969) Recherches sur des mutants bactériens ayant perdu les activités catalytiques liées à la nitrate reductase A. III Caractères biochimiques. Ann Inst Pasteur 117:145–178
Puig J, Azoulay E, Gendre J, Richard E (1969a) Etude génétique des mutants de la région chl A chez Escherichia coli K 12. CR Acad Sci (Paris) 268:183–184
Puig J, Azoulay E, Pichinoty F, Gendre J (1969b) Genetic mapping of the chlC gene of the nitrate reductase A system in Escherichia-coli K 12. Biochem Biophys Res Commun 35:659–662
Rosenfeld SA, Brenchley JE (1980) Bacteriophage P1 as a vehicle for the Mu mutagenesis of Salmonella typhimuricum. J Bacteriol 144:848–851
Ruiz-Herrera J, De Moss JA (1969) Nitrate reductase complex of Escherichia coli K 12. Participation of specific formate dehydrogenase and cytochrome b1 components in nitrate reduction. J Bacteriol 99:720–729
Sanchez Crispin J, Dubourdieu M, Chippaux M (1979) Localization and characterization of cytochromes from membrane vesicles of Escherichia coli K 12 grown in anaerobiosis with nitrate. Biochim Biophys Acta 547:198–210
Spencer ME, Guest JR (1973) Isolation and properties of fumarate reductase mutants of Escherichia coli. J Bacteriol 114:563–570
Venables WA (1972) Genetic studies with nitrate reductase-less mutants of Escherichia coli. I. Fine structure analysis of nar A, nar B and nar E loci. Mol Gen Genet 114:223–231
Author information
Authors and Affiliations
Additional information
Communicated by H. Böhme
Rights and permissions
About this article
Cite this article
Pascal, MC., Chippaux, M. Involvement of a gene of the chl E locus in the regulation of the nitrate reductase operon. Molec. Gen. Genet. 185, 334–338 (1982). https://doi.org/10.1007/BF00330808
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330808