In Vivo Effect of Thiolutin on Cell Growth and Macromolecular Synthesis in
            <i>Escherichia coli</i>

Khachatourians, George G.; Tipper, Donald J.

doi:10.1128/aac.6.3.304

Cited by 27 publications

(19 citation statements)

References 12 publications

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“…The data for holomycin suggest rapid inhibition of RNA chain elongation by the antibiotic and are thus in agreement with the findings of Khachatourians and Tipper (14), who concluded that thiolutin is also an RNA elongation inhibitor. If thiolutin and holomycin inhibit RNA elongation by interaction with RNA polymerase, their binding sites within the enzyme may well differ from that of rifampin, which is an inhibitor of transcriptional initiation (5,11,15,27). This suggestion is consistent with the finding that thiolutin and holomycin are both active against S. aureus strains containing mutations in rpoB that confer resistance to rifampin (20).…”

Section: Discussionsupporting

confidence: 77%

See 1 more Smart Citation

Antimicrobial Properties and Mode of Action of the Pyrrothine Holomycin

Oliva

O’Neill

Wilson

et al. 2001

Antimicrob Agents Chemother

106

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Holomycin, a member of the pyrrothine class of antibiotics, displayed broad-spectrum antibacterial activity, inhibiting a variety of gram-positive and gram-negative bacteria, with the exception of Enterobacter cloacae, Morganella morganii, and Pseudomonas aeruginosa. The antibiotic lacked activity against the eukaryotic microorganisms Saccharomyces cerevisiae and Candida kefyr. Holomycin exhibited a bacteriostatic response against Escherichia coli that was associated with rapid inhibition of RNA synthesis in whole cells. Inhibition of RNA synthesis could have been a secondary consequence of inhibiting tRNA aminoacylation, thereby inducing the stringent response. However, the levels of inhibition of RNA synthesis by holomycin were similar in a stringent and relaxed pair of E. coli strains that were isogenic except for the deletion of the relA gene. This suggests that inhibition of RNA synthesis by holomycin could reflect direct inhibition of DNA-dependent RNA polymerase. Examination of the effects of holomycin on the kinetics of the appearance of ␤-galactosidase in induced E. coli cells was also consistent with inhibition of RNA polymerase at the level of RNA chain elongation. However, holomycin only weakly inhibited E. coli RNA polymerase in assays using synthetic poly(dA-dT) and plasmid templates. Furthermore, inhibition of RNA polymerase was observed only at holomycin concentrations in excess of those required to inhibit the growth of E. coli. It is possible that holomycin is a prodrug, requiring conversion in the cell to an active species that inhibits RNA polymerase.Thiolutin and holomycin (see Fig. 1) are members of the pyrrothine class of naturally occurring antibiotics that are characterized by the possession of a unique pyrrolinonodithiole nucleus (2). Although these antimicrobial agents were originally discovered more than 40 years ago (6, 13, 23, 26), relatively little is known about their mode of action. Limited studies with thiolutin suggest that the pyrrothines may act as inhibitors of DNA-dependent RNA polymerase. Thus, thiolutin preferentially inhibits RNA synthesis in both Saccharomyces cerevisiae (10) and Escherichia coli (14) and is reported to be a potent inhibitor of partially purified RNA polymerases from S. cerevisiae (10, 25). Furthermore, by monitoring the effects of thiolutin on the induction of ␤-galactosidase in E. coli, Khachatourians and Tipper (14) concluded that the antibiotic interfered with RNA chain elongation rather than with initiation of transcription.However, there are several observations that cast doubt upon the hypothesis that thiolutin, and therefore the pyrrothines as a class, prevents microbial growth by interfering with the elongation of RNA transcripts catalyzed by RNA polymerase. For instance, Sivasubramanian and Jayaraman (24) were unable to reproduce the observations made by Khachatourians and Tipper (14) concerning the effects of thiolutin on ␤-galactosidase induction in E. coli. Thus, these authors (24) concluded that thiolutin inhibits initiation of RNA tran...

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Section: Discussionsupporting

confidence: 77%

“…The effects of rifampin and thiolutin on the kinetics of ␤-galactosidase production in E. coli following induction of the lac operon have been used previously to demonstrate that these antibiotics inhibit the initiation and elongation phases, respectively, of mRNA transcription (15).…”

Section: Resultsmentioning

confidence: 99%

Antimicrobial Properties and Mode of Action of the Pyrrothine Holomycin

Oliva

O’Neill

Wilson

et al. 2001

Antimicrob Agents Chemother

106

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“…The most commonly used antibiotic is thiolutin (Jiménez et al,1973), a metabolite produced by Streptomyces luteoreticuli (Celmer and Solomon, 1955) that inhibits all yeast RNA polymerases, mainly at the level of initiation (Tipper, 1973). However, it has also been reported to inhibit elongation in E. coli (Khachatourians and Tipper, 1974). It is commonly used at concentrations around 3 µg/ml but in some cases use of higher concentrations has been reported (Michan et al, 2005;Guan et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

The transcriptional inhibitor thiolutin blocks mRNA degradation in yeast

Pelechano

Pérez-Ortín

2007

Yeast

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Thiolutin is commonly used as a general inhibitor of transcription in yeast. It has been used to calculate mRNA decay rates by stopping the transcription and then determining the relative abundance of individual mRNAs at different times after inhibition. We report here that thiolutin is also an inhibitor of mRNA degradation, and thus its use can lead to miscalculations of mRNA half-lives. The inhibition of mRNA decay seems to affect the mRNA degradation pathway without impeding poly(A) shortening, given that the decay rate of total poly(A) amount is not reduced by thiolutin. Moreover, the thiolutin-dependent inhibition of mRNA degradation has variable effects on different functional groups of genes, suggesting that they use various degradation pathways for their mRNAs.

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“…49 Furthermore, it was also unclear which step of RNA synthesis thiolutin inhibits; results of experiments on beta-galactosidase induction in E. coli have suggested both transcription initiation and chain elongation as potential targets of thiolutin mediated transcription inhibition. 49,50 These contrasting results cast doubt on the proposal that RNA polymerase is the primary target of thiolutin. Nonetheless, these early studies provided valuable insights into thiolutin mode of action, demonstrating for the first time that thiolutin is bacteriostatic and that this effect is reversible.…”

Section: Mode Of Action and Self-resistancementioning

confidence: 99%

Dithiolopyrrolones: biosynthesis, synthesis, and activity of a unique class of disulfide-containing antibiotics

et al. 2014

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Dithiolopyrrolone (DTP) group antibiotics were first isolated in the early half of the 20th century, but only recently has research been reawakened by insights gained from the synthesis and biosynthesis of this structurally intriguing class of molecules. DTPs are characterized by an electronically unique bicyclic structure, which contains a compact disulfide bridge between two ene-thiols. Points of diversity within the compound class occur outside of the bicyclic core, at the two amide nitrogens. Such modifications distinguish three of the most well studied members of the class, holomycin, thiolutin, and aureothricin; the DTP core has also more recently been identified in the marine antibiotic thiomarinol, in which it is linked to a marinolic acid moiety, analog of the FDA-approved topical antibiotic Bactroban® (GlaxoSmithKline). Dithiolopyrrolones exhibit relatively broad-spectrum antibiotic activity against many Gram-positive and Gram-negative bacteria, as well as strains of Mycobacterium tuberculosis. Additionally, they have been shown to exhibit potent and selective anti-cancer activity. Despite this promising profile, there is still much unknown about the mechanisms of action for DTPs. Early reports suggested that they inhibit yeast growth at the level of transcription and that this effect is largely responsible for their distinctive microbial static properties; a similar mechanism is supported in bacteria. Elucidation of biosynthetic pathways for holomycin in Streptomyces clavuligerus and Yersinia ruckeri and thiomarinol in Alteromonas rava sp. nov. SANK 73390, have contributed evidence suggesting that multiple mechanisms may be operative in the activity of these compounds. This review will comprehensively cover the history and development of dithiolopyrrolones with particular emphasis on the biosynthesis, synthesis, biological activity and mechanism of action.

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In Vivo Effect of Thiolutin on Cell Growth and Macromolecular Synthesis in Escherichia coli

Cited by 27 publications

References 12 publications

Antimicrobial Properties and Mode of Action of the Pyrrothine Holomycin

Antimicrobial Properties and Mode of Action of the Pyrrothine Holomycin

The transcriptional inhibitor thiolutin blocks mRNA degradation in yeast

Dithiolopyrrolones: biosynthesis, synthesis, and activity of a unique class of disulfide-containing antibiotics

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