Inhibition of 6-Methylsalicylic Acid Synthesis by the Antibiotic Cerulenin

Ohno, Hiroie; Ohno, Tadao; Awaya, Juichi; Ōmura, Satoshi

doi:10.1093/oxfordjournals.jbchem.a131010

Cited by 37 publications

(18 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fig. 3A shows that cerulenin efficiently inhibits DIF-1 synthesis at concentrations similar to those inhibiting condensing enzymes in other organisms (25,26,44,45).…”

Section: Resultsmentioning

confidence: 71%

See 1 more Smart Citation

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

Kay¹

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

Differentiation-inducing factor (DIF)-1 is a chlorinated alkyl phenone released by developing Dictyostelium amoebae, which induces them to differentiate into stalk cells. A biosynthetic pathway for DIF-1 is proposed from labeling, inhibitor, and enzymological experiments. Cells incorporate 36 Cl؊ into DIF-1 during development, showing that the chlorine atoms originate from chloride ions; peak incorporation is at the first finger stage. DIF-1 synthesis can be blocked by cerulenin, a polyketide synthase inhibitor, suggesting that it is made from a polyketide. This is most likely the C 12 polyketide (2,4,6-trihydroxyphenyl)-1-hexan-1-one (THPH). Feeding experiments confirm that living cells can convert THPH to DIF-1. Conversion requires both chlorination and methylation of THPH, and enzymatic activities able to do this exist in cell lysates. The chlorinating activity, assayed using 36 Cl ؊ , is stimulated by H 2 O 2 and requires both soluble and particulate components. It is specific for THPH and does not use this compound after O-methylation. The methyltransferase is soluble, uses S-adenosyl-L-methionine as a co-substrate, has a K m for dichloro-THPH of about 1.1 M, and strongly prefers this substrate to close analogues. Both chlorinating and methyltransferase activities increase in development in parallel with DIF-1 production, and both are greatly reduced in a mutant strain that makes little DIF-1. It is proposed that DIF-1 is made by the initial assembly of a C 12 polyketide skeleton, which is then chlorinated and methylated.

show abstract

“…Fig. 3A shows that cerulenin efficiently inhibits DIF-1 synthesis at concentrations similar to those inhibiting condensing enzymes in other organisms (25,26,44,45).…”

Section: Resultsmentioning

confidence: 71%

“…The condensing enzymes of polyketide and fatty acid synthases share conserved sequence motifs around the active site (21), and the active site cysteine is normally the target for the covalent inhibitor, cerulenin (22)(23)(24). Inhibition by cerulenin therefore provides a diagnostic test for the proposed polyketide origin of DIF-1 (25,26).…”

Section: Dif-1mentioning

confidence: 99%

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

Kay¹

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Cerulenin has also been found to inhibit the biosynthesis of various polyketide-derived natural products such as 6-methylsalicylic acid [6] and the macrolide antibiotics leucomycin [7] and candicidin [8], suggesting that the condensing enzyme in polyketide biosynthesis resembles the analogous component of fatty acid synthase. This report describes the synthesis of ['Hltetrahydrocerulenin (lb) and its use for specific labelling of fatty acid synthase.…”

mentioning

confidence: 99%

[³H]Tetrahydrocerulenin, a specific reagent for radio‐labelling fatty acid synthases and related enzymes

Roberts

Leadlay

1983

FEBS Letters

View full text Add to dashboard Cite

We have synthesised [3H]tetrahydrocerulenin, a radio‐labelled derivative of cerulenin, an antibiotic from Cephalosporium caerulens which specifically inhibits the condensing enzyme of fatty acid synthases and polyketide synthases from various sources. [3H]Tetrahydrocerulenin binds to fatty acid synthase from pig liver with the same specificity as cerulenin. We have also used [3H]tetrahydrocerulenin to monitor condensing enzyme activity in cell‐free extracts of the erythromycin‐producing organism Streptomyces erythreus.

show abstract

“…Unfortunately there is no available information about the MICs of lincomycin against O. tsutsugamushi. However, according to the MICs of lincomycin against gram-negative bacteria [10], lincomycin is supposed to be much less effective against O. tsutsugamushi because O. tsutsugamushi is one of the gram-negative bacteria. For the example, the MICs of lincomycin against Escherichia coli , one of the typical gram gram-negative bacteria are more than 50 times higher than those against mycoplasmas.…”

Section: Resultsmentioning

confidence: 99%

Decontamination of mycoplasma-contaminated Orientia tsutsugamushi strains by repeating passages through cell cultures with antibiotics

et al. 2013

View full text Add to dashboard Cite

BackgroundMycoplasmas-contamination of Orientia tsutsugamushi, one of the obligated intracellular bacteria, is a very serious problem in in vitro studies using cell cultures because mycoplasmas have significant influence on the results of scientific studies. Only a recommended decontamination method is to passage the contaminated O. tsutsugamushi strains through mice to eliminate only mycoplasmas under influence of their immunity. However, this method sometimes does not work especially for low virulent strains of O. tsutsugamushi which are difficult to propagate in mice. In this study, we tried to eliminate mycoplasmas contaminants from both high virulent and low virulent strains of the contaminated O. tsutsugamushi by repeating passage through cell cultures with antibiotics in vitro.ResultsWe cultured a contaminated, high virulent strain of O. tsutsugamushi using a mouse lung fibroblasts cell line, L-929 cell in the culture medium containing lincomycin at various concentrations and repeated passages about every seven days. At the passage 5 only with 10 μg/ml of lincomycin, we did not detect mycoplasmas by two PCR based methods whereas O. tsutsugamushi continued good growth. During following four passages without lincomycin, mycoplasmas did not recover. These results suggested that mycoplasmas were completely eliminated from the high virulent strain of O. tsutsugamushi. Furthermore, by the same procedures with 10 μg/ml of lincomycin, we also eliminated mycoplasmas from a contaminated, low virulent strain of O. tsutsugamushi. Our additional assay showed that 50 μg/ml of lyncomycin did not inhibit the growth of O. tsutsugamushi, although MICs of many mycoplasmas contaminants were less than 6 μg/ml as shown previously.ConclusionOur results showed an alternative method to eliminate mycoplasmas from the contaminated O. tsutsugamushi strains in place of in vivo passage through mice. Especially this notable method works for the decontamination not only from the high virulent strain also from the low virulent strain of O. tsutsugamushi. For further elimination, lincomycin at the limit concentration, which does not inhibit the growth of O. tsutsugamushi, can possibly eliminate most mycoplasmas from contaminated O. tsutsugamushi strains.

show abstract

Inhibition of 6-Methylsalicylic Acid Synthesis by the Antibiotic Cerulenin

Cited by 37 publications

References 0 publications

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

[³H]Tetrahydrocerulenin, a specific reagent for radio‐labelling fatty acid synthases and related enzymes

Decontamination of mycoplasma-contaminated Orientia tsutsugamushi strains by repeating passages through cell cultures with antibiotics

Contact Info

Product

Resources

About

Inhibition of 6-Methylsalicylic Acid Synthesis by the Antibiotic Cerulenin

Cited by 37 publications

References 0 publications

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

The Biosynthesis of Differentiation-Inducing Factor, a Chlorinated Signal Molecule Regulating DictyosteliumDevelopment

[3H]Tetrahydrocerulenin, a specific reagent for radio‐labelling fatty acid synthases and related enzymes

Decontamination of mycoplasma-contaminated Orientia tsutsugamushi strains by repeating passages through cell cultures with antibiotics

Contact Info

Product

Resources

About

[³H]Tetrahydrocerulenin, a specific reagent for radio‐labelling fatty acid synthases and related enzymes