Overproduction of fumarate reductase in Escherichia coli induces a novel intracellular lipid-protein organelle

Weiner, Joël H.; Lemire, Bernard D.; Elmes, MJ; Bradley, R.; Scraba, Douglas G.

doi:10.1128/jb.158.2.590-596.1984

Cited by 98 publications

(36 citation statements)

References 24 publications

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“…It is possible that the overproduction of membrane-bound proteins from multicopy plasmids may be deleterious to host strains or that growth in the presence of pFAS may deplete the cultures of necessary nutrients for efficient growth. It should be noted, however, that this was not observed for overproduction of QFR in the experiments reported here or by others (2,8,34) or for SQR from pFGS (Fig. 2).…”

Section: Resultscontrasting

confidence: 49%

“…It has previously been shown that, in stationary-phase E. coli cells harboring a fumarate reductase-encoding plasmid similar to pH3, QFR production was amplified 20-fold (34). Because of the limited capacity of the inner membrane, the cells produced novel tubular structures rather than accumulating the extra protein in soluble form, or as inclusion bodies.…”

Section: Resultsmentioning

confidence: 99%

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Anaerobic Expression ofEscherichia coliSuccinate Dehydrogenase: Functional Replacement of Fumarate Reductase in the Respiratory Chain during Anaerobic Growth

1998

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Succinate-ubiquinone oxidoreductase (SQR) from Escherichia coli is expressed maximally during aerobic growth, when it catalyzes the oxidation of succinate to fumarate in the tricarboxylic acid cycle and reduces ubiquinone in the membrane. The enzyme is similar in structure and function to fumarate reductase (menaquinol-fumarate oxidoreductase [QFR]), which participates in anaerobic respiration by E. coli. Fumarate reductase, which is proficient in succinate oxidation, is able to functionally replace SQR in aerobic respiration when conditions are used to allow the expression of the frdABCD operon aerobically. SQR has not previously been shown to be capable of supporting anaerobic growth ofE. coli because expression of the enzyme complex is largely repressed by anaerobic conditions. In order to obtain expression of SQR anaerobically, plasmids which utilize the PFRD promoter of the frdABCD operon fused to the sdhCDAB genes to drive expression were constructed. It was found that, under anaerobic growth conditions where fumarate is utilized as the terminal electron acceptor, SQR would function to support anaerobic growth ofE. coli. The levels of amplification of SQR and QFR were similar under anaerobic growth conditions. The catalytic properties of SQR isolated from anaerobically grown cells were measured and found to be identical to those of enzyme produced aerobically. The anaerobic expression of SQR gave a greater yield of enzyme complex than was found in the membrane from aerobically grown cells under the conditions tested. In addition, it was found that anaerobic expression of SQR could saturate the capacity of the membrane for incorporation of enzyme complex. As has been seen with the amplified QFR complex, E. coli accommodates the excess SQR produced by increasing the amount of membrane. The excess membrane was found in tubular structures that could be seen in thin-section electron micrographs.

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Section: Resultscontrasting

confidence: 49%

Section: Resultsmentioning

confidence: 99%

Anaerobic Expression ofEscherichia coliSuccinate Dehydrogenase: Functional Replacement of Fumarate Reductase in the Respiratory Chain during Anaerobic Growth

1998

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“…The induced cells contain cytoplasmic structures not observed in the parent cells, in which the large amount Of D-LDH produced during induction is most likely to be. We did not observe any intracellular lipid-protein organelles of the type which occur upon overproduction of fumarate reductase in E. ccli (41).…”

Section: Resultscontrasting

confidence: 65%

Overproduction and nucleotide sequence of the respiratory D-lactate dehydrogenase of Escherichia coli

Rule¹,

Pratt²,

Chin³

et al. 1985

J Bacteriol

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Recombinant DNA plasmids containing the gene for the membrane-bound D-lactate dehydrogenase (D-LDH) of Escherichia coli linked to the promoter PL from lambda were constructed. After induction, the levels of D-LDH were elevated 3Q0-fold over that of the wild type and amounted to 35 % of the total cellular protein. The nucleotide sequence of the D-LDH gene was determined and shown to agree with the amino acid composition and the amino-terminal sequence of the purified enzyme. Removal of the amino-terminal formyl-Met from D-LDH was not inhibited in cells which contained these high levels of D-LDH.

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“…This was found with PV 2C and 2BC (2, 10) and 3A and 3AB (references 11 and 14 and unpublished observations) as well as an overexpressed 6-kDa protein of tobacco etch potyvirus (43). Cellular proteins with reported membrane-altering properties include cytochrome B5 (4) and 3-hydroxy-3-methylglutaryl coenzyme A reductase (5,25) as well as a 180-kDa ribosome receptor in yeast (59) and ATP synthase and fumarate reductase in E. coli (58,61). The morphologies of the membrane alterations observed in the reports cited above are quite diverse.…”

Section: Interaction Of Viral Proteins With Intracellular Membranesmentioning

confidence: 96%

Poliovirus 2C protein determinants of membrane binding and rearrangements in mammalian cells

Teterina

Gorbalenya

Egger

et al. 1997

J Virol

150

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Poliovirus protein 2C is a 329-amino acid-protein that is essential for viral RNA synthesis and may perform multiple functions. In infected cells, it is associated with virus-specific membrane vesicles. Recombinant 2C protein expressed in transfected cells has been shown to associate with and induce rearrangement of the intracellular membrane network. This study was designed to map the determinants of membrane binding and rearrangement in the 2C protein. Computer-assisted analysis of the protein sequence led to a prediction that the protein folds into a structure composed of three domains. Expression plasmids that encode each or combinations of these predicted domains were used to examine the abilities of the partial protein sequences to associate with intracellular membranes and to induce rearrangement of these membranes in HeLa cells. Biochemical fractionation procedures suggested that the N-terminal region of the protein was required for membrane association. Electron microscopic and immunoelectron microscopic observation showed that both the N-and C-terminal regions, but not the central portion, of 2C protein interact with intracellular membranes and induce major changes in their morphology. The central portion, when fused to the N-terminal region, altered the specific membrane architecture induced by the N-terminal region, giving rise to vesicles resembling those observed during poliovirus infection.

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Overproduction of fumarate reductase in Escherichia coli induces a novel intracellular lipid-protein organelle

Cited by 98 publications

References 24 publications

Anaerobic Expression ofEscherichia coliSuccinate Dehydrogenase: Functional Replacement of Fumarate Reductase in the Respiratory Chain during Anaerobic Growth

Anaerobic Expression ofEscherichia coliSuccinate Dehydrogenase: Functional Replacement of Fumarate Reductase in the Respiratory Chain during Anaerobic Growth

Overproduction and nucleotide sequence of the respiratory D-lactate dehydrogenase of Escherichia coli

Poliovirus 2C protein determinants of membrane binding and rearrangements in mammalian cells

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