Complete nucleotide sequence of the<i>Escherichia coli</i>gene encoding malate dehydrogenase

McAlister-Henn, Lee; Blaber, Michael; Bradshaw, Ralph A.; Nisco, Steven J.

doi:10.1093/nar/15.12.4993

Cited by 47 publications

(18 citation statements)

References 2 publications

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“…Overall, 49% of the residues in MDH1 are identical with aligned residues in MDH2. MDH1 shares a similar 48% residue identity with E. coli malate dehydrogenase (33) (20,26,33,48) and unlike porcine and murine cytosolic isozymes, which each contain four tryptophan residues (25).…”

Section: Resultsmentioning

confidence: 99%

Isolation, nucleotide sequence analysis, and disruption of the MDH2 gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast malate dehydrogenase.

Minard¹,

McAlister-Henn²

1991

Mol. Cell. Biol.

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The major nonmitochondrial isozyme of malate dehydrogenase (MDH2) in Saccharomyces cerevisiae cells grown with acetate as a carbon source was purified and shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to have a subunit molecular weight of approximately 42,000. Enzyme assays and an antiserum prepared against the purified protein were used to screen a collection of acetate-nonutilizing (acetate-) yeast mutants, resulting in identification of mutants in one complementation group that lack active or immunoreactive MDH2. Transformation and complementation of the acetate-growth phenotype was used to isolate a plasmid carrying the MDH2 gene from a yeast genomic DNA library. The amino acid sequence derived from complete nucleotide sequence analysis of the isolated gene was found to be extremely similar (49% residue identity) to that of yeast mitochondrial malate dehydrogenase (molecular weight, 33,500) despite the difference in sizes of the two proteins. Disruption of the MDH2 gene in a haploid yeast strain produced a mutant unable to grow on minimal medium with acetate or ethanol as a carbon source. Disruption of the MDH2 gene in a haploid strain also containing a disruption in the chromosomal MDHI gene encoding the mitochondrial isozyme produced a strain unable to grow with acetate but capable of growth on rich medium with glycerol as a carbon source. The detection of residual malate dehydrogenase activity in the latter strain confirmed the existence of at least three isozymes in yeast cells.Two isozymes of malate dehydrogenase in mammalian cells catalyze the interconversion of malate plus NAD+ and oxaloacetate plus NADH. The mitochondrial isozyme functions in the tricarboxylic acid cycle, and the cytosolic isozyme catalyzes a step in gluconeogenesis from pyruvate. Also, as components of the malate/aspartate shuttle cycle, the malate dehydrogenase isozymes control the exchange of metabolic intermediates and reducing equivalents across the mitochondrial membrane. Analyses of amino acid sequences for the isozymes from porcine heart (4, 5) and of nucleotide sequences for the porcine and murine cDNAs (25,26) (MATo leu2 his3-AJ trpl gcrl-J-J; 34) was used for isolation of the nonmitochondrial isozyme of malate dehydrogenase. Yeast strains S173-6B (MATa leu2-3,112 his3-AJ ura3-57 trpl-289; 7) and SAMDH1, a derivative of S173-6B containing a chromosomal disruption of the MDH1 gene (47), were used for transformation, growth studies, and cell fractionation. The parental S. cerevisiae strain MMY011 (MATa ade2-1 his3-11,15 leu2-3,112 trpl-l ura3-1 canl-100) and 370 on April 28, 2019 by guest

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Section: Resultsmentioning

confidence: 99%

Isolation, nucleotide sequence analysis, and disruption of the MDH2 gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast malate dehydrogenase.

Minard¹,

McAlister-Henn²

1991

Mol. Cell. Biol.

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“…The sequences of both the mouse and yeast c-and mMDH genes have been reported (Minard and McAlister-Henn, 1991;Setoyama et al, 1988;Takeshima et al, 1988;Thompson et al, 1988). MDH genes have also been isolated from several bacterial sources (McAlister-Henn et al, 1987;Nicholls et al, 1990;Nishiyama et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

Alfalfa malate dehydrogenase (MDH): molecular cloning and characterization of five different forms reveals a unique nodule‐enhanced MDH

Miller¹,

Driscoll²,

Gregerson³

et al. 1998

The Plant Journal

152

100

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SummaryMalate dehydrogenase (MDH) catalyzes the readily reversible reaction of oxaloacetate ≤ malate using either NADH or NADPH as a reductant. In plants, the enzyme is important in providing malate for C 4 metabolism, pH balance, stomatal and pulvinal movement, respiration, β-oxidation of fatty acids, and legume root nodule functioning. Due to its diverse roles the enzyme occurs as numerous isozymes in various organelles. While antibodies have been produced and cDNAs characterized for plant mitochondrial, glyoxysomal, and chloroplast forms of MDH, little is known of other forms. Here we report the cloning and characterization of cDNAs encoding five different forms of alfalfa MDH, including a plant cytosolic MDH (cMDH) and a unique novel nodule-enhanced MDH (neMDH). Phylogenetic analyses show that neMDH is related to mitochondrial and glyoxysomal MDHs, but diverge from these forms early in land plant evolution. Four of the five forms could effectively complement an E. coli Mdh -mutant. RNA and protein blots show that neMDH is most highly expressed in effective root nodules. Immunoprecipitation experiments show that antibodies produced to cMDH and neMDH are immunologically distinct and that the neMDH form comprises the major form of total MDH activity and protein in root nodules. Kinetic analysis showed that neMDH has a turnover rate and specificity constant that can account for the extraordinarily high synthesis of malate in nodules.

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“…This pathway is considered to operate predominantly under anaerobic cell growth conditions (15). However, the effects of different environmental conditions on the regulation of mdh gene expression in E. coli or in other microorganisms have not been examined.The mdh gene in E. coli, which encodes MDH, is located at 70.6 min on the chromosome (8), and it has recently been cloned and sequenced (11,22). The E. coli enzyme is a dimeric enzyme with a molecular weight of 60,000 (13), like the MDHs isolated from most other organisms (14).…”

mentioning

confidence: 99%

“…The mdh gene in E. coli, which encodes MDH, is located at 70.6 min on the chromosome (8), and it has recently been cloned and sequenced (11,22). The E. coli enzyme is a dimeric enzyme with a molecular weight of 60,000 (13), like the MDHs isolated from most other organisms (14).…”

mentioning

confidence: 99%

Regulation of malate dehydrogenase (mdh) gene expression in Escherichia coli in response to oxygen, carbon, and heme availability

1995

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Malate dehydrogenase catalyzes the interconversion of malate and oxaloacetate. It participates as a member of the tricarboxylic acid cycle and the branched noncyclic pathways under aerobic and anaerobic cell growth conditions, respectively. To investigate how the mdh gene is expressed under these different conditions, an mdh-lacZ operon fusion was constructed and analyzed in vivo. The mdh-lacZ fusion was expressed about twofold higher under aerobic conditions than under anaerobic cell growth conditions on most media tested. This anaerobic response is modulated by the ArcA protein, which functions as a repressor of mdh gene expression under both aerobic and anaerobic conditions. In contrast, mutations in the fnr gene did not affect mdh gene expression. Interestingly, cells grown anaerobically with glycerol and trimethylamine N-oxide or fumarate showed higher levels of mdh expression than did cells that were grown aerobically. Depending on the type of carbon compound used for cell growth, mdh expression varied by 11-fold and 5-fold under aerobic and anaerobic conditions, respectively. While mdh transcription was shown to be inversely proportional to the cell growth rate, cellular heme limitation stimulated a fivefold increase in mdh gene expression. The mdh gene appears to be highly regulated to adapt to changing conditions of aerobic and anaerobic cell growth with various types of carbon substrates.Malate dehydrogenase (EC 1.1.1.37) (MDH) is an essential enzyme in the tricarboxylic acid (TCA) cycle as well as the noncyclic anaplerotic pathway of Escherichia coli (15). It catalyzes the interconversion of malate and oxaloacetate with a concomitant reduction of NAD or oxidation of NADH. The reaction carried out by MDH under aerobic conditions in which the TCA cycle is operative is from malate to oxaloacetate (i.e., oxidation); the reaction proceeds in the opposite direction for participation in the branched noncyclic pathway. This pathway is considered to operate predominantly under anaerobic cell growth conditions (15). However, the effects of different environmental conditions on the regulation of mdh gene expression in E. coli or in other microorganisms have not been examined.The mdh gene in E. coli, which encodes MDH, is located at 70.6 min on the chromosome (8), and it has recently been cloned and sequenced (11,22). The E. coli enzyme is a dimeric enzyme with a molecular weight of 60,000 (13), like the MDHs isolated from most other organisms (14). It has a remarkable homology to the mitochondrial MDH (e.g., 59% amino acid identity to porcine mitochondrial MDH), especially in the nucleotide binding site located near the amino terminus of the protein. The E. coli enzyme shows a lower degree of sequence similarity to the alternative eucaryotic MDH isozyme that is cytoplasmically located (ca. 20% similarity to the porcine cytoplasmic MDH). The level of the E. coli MDH enzyme in the cell is reduced by anaerobiosis and glucose (6, 21) and induced by acetate and malate (21). In addition, MDH activity is elevated in ar...

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Complete nucleotide sequence of theEscherichia coligene encoding malate dehydrogenase

Cited by 47 publications

References 2 publications

Isolation, nucleotide sequence analysis, and disruption of the MDH2 gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast malate dehydrogenase.

Isolation, nucleotide sequence analysis, and disruption of the MDH2 gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast malate dehydrogenase.

Alfalfa malate dehydrogenase (MDH): molecular cloning and characterization of five different forms reveals a unique nodule‐enhanced MDH

Regulation of malate dehydrogenase (mdh) gene expression in Escherichia coli in response to oxygen, carbon, and heme availability

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