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Metabolic and Transcriptional Response to Cofactor Perturbations in Escherichia coli

Holm, A. K. (author)
Blank, L. M. (author)
Oldiges, M. (author)
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Schmid, A. (author)
Solem, C. (author)
Jensen, P. R. (author)
Vemuri, Goutham, 1976 (author)
Chalmers tekniska högskola,Chalmers University of Technology
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 (creator_code:org_t)
2010
2010
English.
In: Journal of Biological Chemistry. - 0021-9258 .- 1083-351X. ; 285:23, s. 17498-17506
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • Metabolic cofactors such as NADH and ATP play important roles in a large number of cellular reactions, and it is of great interest to dissect the role of these cofactors in different aspects of metabolism. Toward this goal, we overexpressed NADH oxidase and the soluble F1-ATPase in Escherichia coli to lower the level of NADH and ATP, respectively. We used a global interaction network, comprising of protein interactions, transcriptional regulation, and metabolic networks, to integrate data from transcription profiles, metabolic fluxes, and the metabolite levels. We identified high-scoring networks for the two strains. The results revealed a smaller, but denser network for perturbations of ATP level, compared with that of NADH level. The action of many global transcription factors such as ArcA, Fnr, CRP, and IHF commonly involved both NADH and ATP, whereas others responded to either ATP or NADH. Overexpressing NADH oxidase invokes response in widespread aspects of metabolism involving the redox cofactors (NADH and NADPH), whereas ATPase has a more focused response to restore ATP level by enhancing proton translocation mechanisms and repressing biosynthesis. Interestingly, NADPH played a key role in restoring redox homeostasis through the concerted activity of isocitrate dehydrogenase and UdhA transhydrogenase. We present a reconciled network of regulation that illustrates the overlapping and distinct aspects of metabolism controlled by NADH and ATP. Our study contributes to the general understanding of redox and energy metabolism and should help in developing metabolic engineering strategies in E. coli.

Subject headings

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology (hsv//eng)

Keyword

protein
topology
flux
Chromatography-mass spectrometry
promoters
cloning
gene-expression
k-12
network
nadh availability

Publication and Content Type

art (subject category)
ref (subject category)

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