2018
DOI: 10.1002/anie.201807970
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Genome Editing Reveals Novel Thiotemplated Assembly of Polythioamide Antibiotics in Anaerobic Bacteria

Abstract: Closthioamide (CTA) is a unique symmetric nonribosomal peptide with six thioamide moieties that is produced by the Gram-positive obligate anaerobe Ruminiclostridium cellulolyticum. CTA displays potent inhibitory activity against important clinical pathogens, making it a promising drug candidate. Yet, the biosynthesis of this DNA gyrase-targeting antibiotic has remained enigmatic. Using a combination of genome mining, genome editing (targeted group II intron, CRISPR/Cas9), and heterologous expression, we show t… Show more

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Cited by 34 publications
(51 citation statements)
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“…8Ac), thereby decrypting the previously unknown origins of the thioamide moieties present in other peptides (for example, thiopeptin and Sch 18640) 132 and consistent with the involvement of YcaO-TfuA-like protein pairs in the biosynthesis of thioviridamide (27)-like RiPPs [133][134][135][136][137] . Thioamide bonds are also present in a number of other non-RiPP NPs, such as closthioamide (28), a remarkable symmetrical peptide comprising six thioamide moieties 138,139 , and 6thioguanine (29), a guanosine nucleotide mimic with potent DNA-targeting and RNAtargeting cytotoxicity 140,141 . In these examples however, thioamide formation is catalysed by members of a novel family of alpha hydrolase (AANH)-like proteins and not YcaO proteins 139,141 .…”
Section: Ycao Proteinsmentioning
confidence: 99%
See 1 more Smart Citation
“…8Ac), thereby decrypting the previously unknown origins of the thioamide moieties present in other peptides (for example, thiopeptin and Sch 18640) 132 and consistent with the involvement of YcaO-TfuA-like protein pairs in the biosynthesis of thioviridamide (27)-like RiPPs [133][134][135][136][137] . Thioamide bonds are also present in a number of other non-RiPP NPs, such as closthioamide (28), a remarkable symmetrical peptide comprising six thioamide moieties 138,139 , and 6thioguanine (29), a guanosine nucleotide mimic with potent DNA-targeting and RNAtargeting cytotoxicity 140,141 . In these examples however, thioamide formation is catalysed by members of a novel family of alpha hydrolase (AANH)-like proteins and not YcaO proteins 139,141 .…”
Section: Ycao Proteinsmentioning
confidence: 99%
“…Thioamide bonds are also present in a number of other non-RiPP NPs, such as closthioamide (28), a remarkable symmetrical peptide comprising six thioamide moieties 138,139 , and 6thioguanine (29), a guanosine nucleotide mimic with potent DNA-targeting and RNAtargeting cytotoxicity 140,141 . In these examples however, thioamide formation is catalysed by members of a novel family of alpha hydrolase (AANH)-like proteins and not YcaO proteins 139,141 .…”
Section: Ycao Proteinsmentioning
confidence: 99%
“…As before, no oxygen isosteres of known CTA congeners were detected in culture extracts (see Figure S3). We next turned to an Escherichia coli heterologous expression system for the production of polythioamidated peptides . A vector containing all of the CTA biosynthetic genes except ctaC was constructed (pCTA‐ ΔctaC ; see Figure S4) and the metabolite profile of E. coli pCTA‐ ΔctaC was checked for the accumulation of intermediates.…”
Section: Figurementioning
confidence: 99%
“…However, it is possible that the type II intron inserted in ctaC causes polar effects . We therefore used a CRISPR‐Cas9 genome editing method to generate a new R. cellulolyticum ΔctaC strain with a nonsense mutation in ctaC (see Figure S2) . As before, no oxygen isosteres of known CTA congeners were detected in culture extracts (see Figure S3).…”
Section: Figurementioning
confidence: 99%