Nucleotide sequence of<i>metF</i>, the<i>E. coli</i>structural gene for 5–10 methylene tetrahydrofolate reductase and of its control region

Saint‐Girons, Isabelle; Duchange, Nathalie; Zakin, Mario M.; Park, Insoo; Margarita, Danielle; Ferrara, Pascual; Cohen, Georges N.

doi:10.1093/nar/11.19.6723

Cited by 61 publications

(33 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The function of the C-terminal domain of plant MTHFRs, which are insensitive to AdoMet, is unknown (9). The NADH-dependent, AdoMet-insensitive MTHFR of Escherichia coli (MetF) lacks the C-terminal domain (15)(16)(17).…”

mentioning

confidence: 99%

Metabolic Engineering in Yeast Demonstrates ThatS-Adenosylmethionine Controls Flux through the Methylenetetrahydrofolate Reductase Reaction in Vivo

Roje¹,

Chan²,

Kaplan³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

One-carbon flux into methionine and S-adenosylmethionine (AdoMet) is thought to be controlled at the methylenetetrahydrofolate reductase (MTHFR) step. Mammalian MTHFRs are inhibited by AdoMet in vitro, and it has been proposed that methyl group biogenesis is regulated in vivo by this feedback loop. In this work, we used metabolic engineering in the yeast Saccharomyces cerevisiae to test this hypothesis. Like mammalian MTHFRs, the yeast MTHFR encoded by the MET13 gene is NADPH-dependent and is inhibited by AdoMet in vitro.ThiscontrastswithplantMTHFRs,whichareNADHdependent and AdoMet-insensitive. To manipulate flux through the MTHFR reaction in yeast, the chromosomal copy of MET13 was replaced by an Arabidopsis MTHFR cDNA (AtMTHFR-1) or by a chimeric sequence (Chimera-1) comprising the yeast N-terminal domain and the AtMTHFR-1 C-terminal domain. Chimera-1 used both NADH and NADPH and was insensitive to AdoMet, supporting the view that the C-terminal domain is responsible for AdoMet inhibition. Engineered yeast expressing Chimera-1 accumulated 140-fold more AdoMet and 7-fold more methionine than did the wild-type and grew normally. Yeast expressing AtMTHFR-1 accumulated 8-fold more AdoMet. This is the first in vivo evidence that the AdoMet sensitivity and pyridine nucleotide preference of MTHFR control methylneogenesis.13 C labeling data indicated that glycine cleavage becomes a more prominent source of one-carbon units when Chimera-1 is expressed. Possibly related to this shift in one-carbon fluxes, total folate levels are doubled in yeast cells expressing Chimera-1.

show abstract

mentioning

confidence: 99%

Metabolic Engineering in Yeast Demonstrates ThatS-Adenosylmethionine Controls Flux through the Methylenetetrahydrofolate Reductase Reaction in Vivo

Roje¹,

Chan²,

Kaplan³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…We decided in 1981 to study the structure and regulation of the enzymes leading to methionine from homoserine, and with Mario Zakin, Claude Parsot, Nathalie Duchange, Isabelle Saint Girons, and Alice Dautry, we determined the primary structure of aspartokinase II-homoserine dehydrogenase II, cystathionine-γ -synthase, β-cystathionase, and methylene tetrahydrofolate reductase, as well as that of the methionine repressor we had mentioned in our 1953 paper (28,47,48,56). All of the methionine genes, with the exception of metH and metG, are subject to repression by methionine and the repression is mediated through the MetJ gene product, a common aporepressor.…”

Section: Back To the Institut Pasteur (1969-1989): The Sequence Of Thmentioning

confidence: 99%

“…It was reasonable to assume that the repressor binding sites should be similar for all genes regulated via repression by methionine. Preliminary studies had shown us that upstream two of the structural genes, homologous sequences could be operator structures recognized by the same regulatory protein (48). Further work showed us that the repressor binding site is composed of an 8-nucleotide-long unit (3).…”

Section: Back To the Institut Pasteur (1969-1989): The Sequence Of Thmentioning

confidence: 99%

Looking Back

Cohen

2005

Annu. Rev. Microbiol.

Self Cite

View full text Add to dashboard Cite

My encounter with Jacques Monod has shaped my scientific career. After a short incursion in the biochemistry of strict anaerobes, and after elucidating the biosynthetic pathway leading from aspartate to threonine in Escherichia coli, I joined his laboratory. With him and Howard Rickenberg, I discovered the stereospecific permeability of galactosides and amino acids (permeases). After this intermezzo, I returned to the analysis of biosynthetic pathways and of their regulation by allosteric feedback inhibition and repression in E. coli. Among others, my studies led to the discovery of the tryptophan and methionine repressors, to the incorporation of amino acid analogues in proteins, including selenomethionine (which much later led to progress in protein crystallography), to the definition of isofunctional and multifunctional enzymes, and to the elucidation of the primary structure of most of the enzymes leading to threonine and methionine.

show abstract

“…The pertinent DNA sequences of the metA, metC, metF, and metJ/metBL promoter regions contain from two to five imperfect, tandem repeats of the Met-box consensus sequence (3,4,7,18,24,25). The homology of individual Met-box sequences varies significantly within and between met regulon elements (3, 4).…”

mentioning

confidence: 99%

The Escherichia coli K-12 metJ193 allele contains a point mutation which alters the hydrophobic pocket responsible for in vitro binding of S-adenosylmethionine: effects on cell growth and induction of met regulon expression

Collier

Johnson

1990

J Bacteriol

View full text Add to dashboard Cite

The metJ193 allele encodes one of two identified temperature-sensitive Escherichia coli K-12 (34C). When 28°C cultures of strains bearing two metJl93 aDleles were transferred from methionine-containing medium to minimal medium, derepression of met regulon expression did not occur quickly enough to avoid a lag in growth due to the methionine deprivation. The inability of the MetJ193 protein to easily accomplish transition between apo-and active-repressor conformations was also demonstrated by using a maxicell system to study expression of a plasmid-borne copy of the E. coli metF transcription unit. These results confirm the importance of the leucine 56 residue for the structure and function in vivo of the wild-type MetJ protein.

show abstract

Nucleotide sequence ofmetF, theE. colistructural gene for 5–10 methylene tetrahydrofolate reductase and of its control region

Abstract: The nucleotide sequence of the E.coli metF gene (888 nucleotides), coding for 5-10 methylene tetrahydrofolate reductase, has been determined.

Cited by 61 publications

References 19 publications

Metabolic Engineering in Yeast Demonstrates ThatS-Adenosylmethionine Controls Flux through the Methylenetetrahydrofolate Reductase Reaction in Vivo

Metabolic Engineering in Yeast Demonstrates ThatS-Adenosylmethionine Controls Flux through the Methylenetetrahydrofolate Reductase Reaction in Vivo

Looking Back

The Escherichia coli K-12 metJ193 allele contains a point mutation which alters the hydrophobic pocket responsible for in vitro binding of S-adenosylmethionine: effects on cell growth and induction of met regulon expression

Contact Info

Product

Resources

About