The seleno-polypeptide of formic dehydrogenase (formate hydrogen-lyase linked) from Escherichia coli: genetic analysis

Pecher, A.; Zinoni, F; Böck, August

doi:10.1007/bf00428850

Cited by 59 publications

(32 citation statements)

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“…In experiments similar to those presented in Table 2, strain M9s, a known fdhF mutant (24), produced 75 to 85% of the hydrogen uptake activity of parent strain MC4100, measured either as benzyl viologen or fumarate reduction. The HYD activity of this strain, measured as tritium exchange, was close to 100% of the parent level.…”

Section: Resultsmentioning

confidence: 80%

“…The flhB gene is located near the fdhF gene, which maps at 92.4 min (24,42), and the two mutant phenotypes can be readily distinguished by the hydrogen uptake characteristics of the strains. In experiments similar to those presented in Table 2, strain M9s, a known fdhF mutant (24), produced 75 to 85% of the hydrogen uptake activity of parent strain MC4100, measured either as benzyl viologen or fumarate reduction.…”

Section: Resultsmentioning

confidence: 99%

“…Formate is further metabolized by formate hydrogenlyase (FHL) to dihydrogen and carbon dioxide (13). Various components of the FHL complex, a unique formate dehydrogenase isoenzyme (FDH-H) and a specific hydrogenase isoenzyme (HYD3), have been identified, and the genes coding for these proteins (fdhF and hyd-17, respectively) have been described (10,24,32,41,45). These genes are transcribed only under anaerobic conditions and require formate as an inducer (4,25,26,42,44).…”

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Genetic regulation of formate hydrogenlyase of Escherichia coli: role of the fhlA gene product as a transcriptional activator for a new regulatory gene, fhlB

Maupin

Shanmugam

1990

J Bacteriol

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A new gene whose product is required for the production of formate hydrogenlyase (FHL) has been identified in Escherichia coli. This gene, termedjhlB, maps between thefrdA (94.4 min) and argl (96.6 min) genes on the E. coli chromosome and is transcribed in a clockwise direction toward argI. Biochemical analysis of an FhIBmutant, strain SE-2011 [(I(fldB-lacZ+)], revealed that the mutant lacks formate dehydrogenase activity associated with FHL (FDH-H) and hydrogenase activity. As a result of these defects, fermentative hydrogen production and hydrogen uptake reactions were undetectable in strain SE-2011. Fumarate reductase activity of this mutant was also reduced to about 15% of the levels of the parent (strain MC4100), and strain SE-2011 did not produce succinate as a fermentation end product. Regulation of expression of theJflB gene, studied as production of ,-galactosidase activity by strain SE-2011, revealed that the operon is expressed at low levels under aerobic conditions. Under anaerobic growth conditions, this activity increased by two-to threefold. Addition of formate enhanced the differential rate of synthesis of thefluB gene product to as high as 130 U of ,B-galactosidase specific activity per ,ug of cell protein, but only under anaerobic conditions. Formatedependent expression of I(ffllB-lacZ+) required the ar-subunit of RNA polymerase and theflUA gene product. Under anaerobic growth conditions, Escherichia coli ferments glucose to yield acetate, ethanol, succinate, lactate, and formate (9). Formate is further metabolized by formate hydrogenlyase (FHL) to dihydrogen and carbon dioxide (13). Various components of the FHL complex, a unique formate dehydrogenase isoenzyme (FDH-H) and a specific hydrogenase isoenzyme (HYD3), have been identified, and the genes coding for these proteins (fdhF and hyd-17, respectively) have been described (10,24,32,41,45). These genes are transcribed only under anaerobic conditions and require formate as an inducer (4,25,26,42,44). Several other genes whose products are essential for production of one or both of the enzyme activities have also been described. These include the hyd, fllA, chl, mol, and rpoN genes (3, 6, 18, 23, 25, 28-31, 33, 37, 38, 40, 43). Among these genes, mutations in the fhlA and rpoN genes affect only the production of FHL' activity, and these gene products are needed for transcription of the fdhF and hyd-17 genes (3,28,29,33). This report presents evidence of an additional gene, ffhlB, which is also needed for production of FHL activity of E. coli. Transcription of this gene requires the FhIA protein.The role of the FhlA protein as a putative transcriptional activator in regulation of theflzlB gene was deduced from the * Corresponding author.

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Section: Resultsmentioning

confidence: 80%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Genetic regulation of formate hydrogenlyase of Escherichia coli: role of the fhlA gene product as a transcriptional activator for a new regulatory gene, fhlB

Maupin

Shanmugam

1990

J Bacteriol

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“…The FDH-H polypeptide is encoded by the fdhF gene at 92 min on the E. coli map (27). The FdnG and FdhF polypeptides each contain selenium in the form of selenocysteine, and each associates with molybdenum cofactor.…”

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Genetic evidence that genes fdhD and fdhE do not control synthesis of formate dehydrogenase-N in Escherichia coli K-12

1991

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Enterobacteria synthesize two formate dehydrogenases, formate dehydrogenase-N (encoded byfdnGHI) and formate dehydrogenase H (encoded by fdhF). Previous work has identified two rha-linked Salmonella typhimurium genes, fdnB and fdnC, which are required primarily for formate dehydrogenase-N activity. Analogous mutants, termed fdhD and fdhE, have been isolated in Escherichia coli. We used gene fusions betweenfdnG, the structural gene for the large subunit of formate dehydrogenase-N, and lacZ, the structural gene for I-galactosidase, to examine E. coli fdnGHI operon expression in fdhD and fdhE insertion mutants. Expression of the 4'(fdnG-lacZ) gene fusions was little affected by these insertions, suggesting that fdhD and fdhE do not control transcription or UGA decoding of the formate dehydrogenase-N structural genes. Our complementation tests, with cloned E. coli fdhD and fdhE genes, indicate that the S. typhimurium fdnC and fdnB genes are functionally homologous to the E. coli fdhD and fdhE genes, respectively.Formate (HCO2-) is a product of anaerobic pyruvate cleavage. Enterobacteria have two pathways for anaerobic metabolism of formate (Fig. 1). A respiratory pathway, formate-nitrate oxidoreductase, allows for energy conservation via oxidative phosphorylation during anaerobic growth. This pathway involves two membrane-bound, multisubunit enzymes, formate dehydrogenase-N (FDH-N) and respiratory nitrate reductase. Synthesis of this enzyme complex is induced by nitrate during anaerobic growth. FDH-N and respiratory nitrate reductase are associated with specific cytochromes, cyt bId' and cyt bNa,j, respectively. The strongly electronegative redox potential of the C02-HCO2-couple (Eo' = -432 mV) makes formate an efficient electron donor for respiratory nitrate reduction. The second pathway, formate-hydrogen lyase, operates anaerobically in the absence of nitrate. This pathway involves two enzymes, formate dehydrogenase-H (FDH-H) and hydrogenase 3. Synthesis of this enzyme complex is induced by anaerobiosis only in the absence of nitrate. The exact physiological role of formate-hydrogen lyase is not established. The H2 and CO2 produced by formate-hydrogen lyase accumulates in Durham tubes, so gas production provides a qualitative estimate of this activity. Formate metabolism in enterobacteria has been recently reviewed (33).The three subunits of FDH-N are encoded by the fdnGHI operon at 32 min on the Escherichia coli genetic map (5). The FDH-H polypeptide is encoded by the fdhF gene at 92 min on the E. coli map (27). The FdnG and FdhF polypeptides each contain selenium in the form of selenocysteine, and each associates with molybdenum cofactor. Thus, mutants defective in selenocysteine (sel) or molybdenum cofactor (chl) synthesis or incorporation are devoid of formate dehydrogenase activity (for reviews, see references 7 and 33).Analysis of E. coli and Salmonella typhimurium has identified four sel genes (originally termed fdh), lesions in which abolish the activities of 20,24). The selA (80 min) and selD (38 min) gen...

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“…The first step of this pathway involves the product of the structural genefdhF. It is located at 93 min on the E. coli linkage map (Pecher et al, 1985) and is transcribed counterclockwise on the chromosome (Wu & Mandrand-Berthelot, 1987). Its expression is induced by formate and repressed by nitrate and trimethylamine N-oxide.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Product of the Cloned fdhF Gene of Escherichia coli

Mandrand‐Berthelot²

1987

Microbiology

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The seleno-polypeptide of formic dehydrogenase (formate hydrogen-lyase linked) from Escherichia coli: genetic analysis

Cited by 59 publications

References 32 publications

Genetic regulation of formate hydrogenlyase of Escherichia coli: role of the fhlA gene product as a transcriptional activator for a new regulatory gene, fhlB

Genetic regulation of formate hydrogenlyase of Escherichia coli: role of the fhlA gene product as a transcriptional activator for a new regulatory gene, fhlB

Genetic evidence that genes fdhD and fdhE do not control synthesis of formate dehydrogenase-N in Escherichia coli K-12

Characterization of the Product of the Cloned fdhF Gene of Escherichia coli

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