Biology subjects

Rigali, S.

Publications and source records attributed to Rigali, S..

At least 19 recordsLinked to original sources

bioRxiv2026

Bacillus adaptation to Pseudomonas secondary metabolites enhances its root competitiveness

Bacillus velezensis is a widely used plant growth-promoting rhizobacterium whose effectiveness under natural conditions is strongly influenced by interactions with surrounding microorganisms. While bacterial secondary metabolites are known to shape these interactions, little is known about their long-term evolutionary consequences. Here, we show that repeated exposure of B. velezensis GA1 to secondary metabolites produced by the competing rhizobacterium Pseudomonas sessilinigenes CMR12a drives the emergence of an adapted subpopulation with enhanced ecological fitness. Multi-omics analyses revealed extensive metabolomic and transcriptional changes associated with altered growth dynamics, sporulation, motility, and biofilm formation. Importantly, the evolved variant exhibited improved tomato root colonization and reduced the abundance of the competing Pseudomonas strain in planta. Together, our results demonstrate that prolonged exposure to diffusible bacterial metabolites can drive rapid adaptive diversification in rhizosphere-associated bacteria and highlight the importance of long-term interbacterial interactions in shaping the outcome of plant microbiome assembly and biocontrol performance.

microbiology↗

bioRxiv2026

A global regulatory atlas of Streptomyces reveals conserved and diversified transcriptional networks across actinomycetes

Transcriptional regulatory networks determine how bacteria integrate environmental signals with growth, metabolism and stress adaptation. Actinomycetes encode exceptionally large repertoires of transcription factors (TFs) coordinating morphological development, environmental adaptation and specialized metabolism, yet their regulatory networks remain poorly defined. Here, we apply DNA affinity purification sequencing (DAP-seq) to 789 predicted TFs of Streptomyces coelicolor, generating genome-wide binding maps for 393 regulators and expanding the experimentally supported regulome from [~]8% to [~]50%. Integration with ChIP-seq reveals pleiotropic regulators and hierarchical network architecture linking primary metabolism, development and biosynthetic gene clusters (BGCs). Multiplexed DAP-seq (multiDAP) across 16 additional actinomycetes uncovered deeply conserved regulatory circuits alongside widespread divergence in TF-target interactions. Together, these data establish a regulatory atlas for Streptomyces and related actinomycetes, enabling researchers to explore control of genes, pathways, BGCs and conserved network modules. This provides a foundation for predictive analysis and engineering of complex bacterial phenotypes in biotechnology and medicine.

microbiology↗

bioRxiv2025

The antifungal and other bioactive properties of the volatilome of Streptomyces scabiei

Volatile compounds (VCs) produced by most host-associated bacteria remain largely unexplored despite their potential roles in suppressing microbial competitors and facilitating host colonization. This study investigated the volatilome of Streptomyces scabiei 87-22, the model species for causative agents of common scab in root and tuber crops, under culture conditions that completely inhibited fungal growth, including the phytopathogens Alternaria solani and Gibberella zeae. Bicameral assays confirmed that these inhibitory effects were at least partially mediated by VCs. Using GC x GC TOFMS, 36 VCs were unambiguously identified as products of S. scabiei 87-22 metabolic activity. These included mainly ketones and aromatic compounds (both benzene derivatives and heterocycles), along with smaller contributions from other chemical families including sulfur-containing compounds, nitriles, esters, terpenoids, an amide, and an aldehyde. A literature survey suggests that many of these VCs possess antibacterial, antifungal, anti-oomycete, nematocidal, and insecticidal effects, while the bioactivity of others remains speculative, having been identified only within complex volatile mixtures. Among those with known antifungal properties, dimethyl trisulfide, 2-heptanone, and creosol inhibited the growth of the fungal pathogens tested in this study. In addition, we reveal here that 3-penten-2-one is also a strong inhibitor of fungal growth. Remarkably, despite S. scabiei 87-22 being defined as a pathogen, some of its VCs were associated with plant growth promotion and defense stimulation. Overall, our work highlights the remarkable potential of S. scabiei 87-22 to produce VCs with diverse bioactivities including fungal pathogen inhibition and priming plant resistance. ImportanceThis study reveals that Streptomyces scabiei, the bacterium causing common scab in root and tuber crops, produces a wide variety of volatile chemicals with surprising benefits. These natural compounds can inhibit growth of other harmful microbes, including fungal plant pathogens. Some of these chemicals are already known to fight pests and diseases, while others, like 3-penten-2-one, are newly discovered as potential antifungals. Even more unexpectedly, some of the identified compounds may help plants grow or boost their defenses. Combined with previous work, our findings challenge the idea that S. scabiei is purely harmful and suggests it might, under certain conditions, play a protective role in its environment. This work deepens our understanding of how microbes interact with each other and with plants, and could help shape more sustainable approaches to agriculture.

microbiology↗

bioRxiv2025

The Transcriptional Architecture of Bacterial Biosynthetic Gene Clusters

Bacteria produce diverse bioactive metabolites with ecological and pharmaceutical importance. These compounds are synthesized by biosynthetic gene clusters (BGCs), whose expression is tightly regulated. While many studies have examined the factors influencing BGC expression, including transcription factors (TFs) and environmental signals, the regulatory architecture governing BGCs expression remains largely unexplored. In this meta-analysis, we collected experimental datasets of bacterial transcription factor binding sites (TFBSs) to unveil i) the functional gene categories preferentially targeted by TFs, ii) the regulatory coverage based on cluster organization, iii) the positional distribution of TFBSs, and iv) the binding strength of TFs. Our analysis reveals a regulatory strategy where global TFs primarily target pathway-specific TFs when present, aligning with a "one-for-all" strategy ensuring cluster-wide expression control. Additionally, examination of the organization of TFBS-associated genes identified distinct transcriptional strategies: regulatory genes are frequently monocistronic, while biosynthetic genes tend to be co-transcribed in operons to guarantee biosynthesis efficiency. The positional distribution of TFBSs highlights a strong enrichment in the upstream regions of genes optimizing their role in gene regulation. Finally, assessment of TF-TFBS interaction strength suggests that TFBSs within BGCs exhibit lower binding affinities compared to those associated with core regulon genes that reside outside BGCs, allowing greater regulatory flexibility in response to multiple environmental cues. These findings provide new insights into the regulatory principles shaping BGC expression and would help predict conditions for activating cryptic BGCs, facilitating the discovery of novel bioactive compounds through targeted culture and engineering strategies.

microbiology↗

bioRxiv2024

A new pathway in central metabolism mediates nutrient control of development and antibiotic production by Streptomyces

The amino sugar N-acetylglucosamine (GlcNAc) plays a central role in primary metabolism and is a key signaling molecule for the onset of morphological and chemical differentiation of Streptomyces. The global nutrient-sensory regulator DasR acts as the gatekeeper of development in streptomycetes, and its activity is modulated by aminosugar phosphates. Here, we report the discovery of a novel pathway in aminosugar metabolism that governs GlcNAc sensing. GlcNAc-6P is converted into a toxic metabolite via two new enzyme functions, namely dehydration of N-acetylglucosamine-6-phosphate by NagS to form 6P-Chromogen I, a reaction that has not yet been described in the textbooks, and its subsequent deacetylation by NagA producing a cytotoxic structural analogue of ribose. The latter reveals an unexpected promiscuous activity for GlcNAc-6P deacetylase NagA. The crystal structures of NagS apoenzyme and NagS in complex with its substrate GlcNAc-6P or its inhibitor 6-phosphogluconate were resolved at 2.3 [A], 2.6 [A], and 1.7 [A] resolution, respectively. Detailed in vivo and in vitro studies resolved the key residues of the NagS catalytic site. Thus, we have uncovered a novel pathway in aminosugar metabolism that sheds new light on nutrient-mediated control of development and antibiotic production in Streptomyces.

microbiology↗

bioRxiv2024

LogoMotif: a comprehensive database of transcription factor binding site profiles in Actinobacteria

Actinobacteria undergo a complex multicellular life cycle and produce a wide range of specialized metabolites, including the majority of the antibiotics. These biological processes are controlled by intricate regulatory pathways, and to better understand how they are controlled we need to augment our insights into the transcription factor binding sites. Here, we present LogoMotif (https://logomotif.bioinformatics.nl), an open-source database for characterized and predicted transcription factor binding sites in Actinobacteria, along with their cognate position weight matrices and hidden Markov models. Genome-wide predictions of binding site locations in Streptomyces model organisms are supplied and visualized in interactive regulatory networks. In the web interface, users can freely access, download and investigate the underlying data. With this curated collection of actinobacterial regulatory interactions, LogoMotif serves as a basis for binding site predictions, thus providing users with clues on how to elicit the expression of genes of interest and guide genome mining efforts. HighlightsO_LIActinobacterial regulatory networks are key for compound discovery, including antibiotics. C_LIO_LIContains [~]400 validated and [~]12,100 predicted interactions, presented in interactive networks. C_LIO_LIServes as foundation for regulatory predictions in the gene cluster detection tool, antiSMASH. C_LIO_LILogoMotifs data and algorithms provide knowledge on expression and functional inference of genes. C_LIO_LILogoMotif aids in the discovery of novel chemistry within Actinobacteria and beyond. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=74 SRC="FIGDIR/small/582527v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@cfb7cdorg.highwire.dtl.DTLVardef@12c017borg.highwire.dtl.DTLVardef@155096org.highwire.dtl.DTLVardef@15dcbcc_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

bioRxiv2023

Nitrogen sources enhance siderophore-mediated competition for iron between potato common scab and late blight causative agents

Withdrawal StatementThe authors have withdrawn their manuscript owing to the fact that the P. infestans strain received from third parties is not the one referred in the manuscript despite having the certificate from the providing collection. All the experiments and data are correct but not included into a proper research context. A new version of this work will be published in near future. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

microbiology↗

bioRxiv2023

The fate of bacterial secondary metabolites in the rhizosphere: Streptomyces degrades and feeds on cyclic lipopeptides produced by competitors

Cyclic lipopeptides are key bioactive secondary metabolites produced by some plant beneficial rhizobacteria such as Pseudomonas and Bacillus. They exhibit antimicrobial properties, promote induced systemic resistance in plants and support key developmental traits including motility, biofilm formation and root colonization. However, our knowledge about the fate of lipopeptides once released in the environment and especially upon contact with neighboring rhizobacteria remains limited. Here, we investigated the enzymatic degradation of Bacillus and Pseudomonas cyclic lipopeptides by Streptomyces venezuelae. We observed that Streptomyces is able to degrade the three lipopeptides surfactin, iturin and fengycin upon confrontation with of B. velezensis in vitro and in planta according to specific mechanisms. S. venezuelae was also able to degrade the structurally diverse sessilin, tolaasin, orfamide, xantholisin and putisolvin-type lipopeptides produced by Pseudomonas, indicating that this trait is likely engage in the interaction with various competitors.Furthermore, the degradation of CLPs is associated with the release of free amino and fatty acids and was found to enhance Streptomyces growth, indicating a possible nutritional utilization. Thereby, this work stresses on how the enzymatic arsenal of S. venezuelae may contribute to its adaptation to BSMs-driven interactions with microbial competitors. The ability of Streptomyces to degrade exogenous lipopeptides and feed on them adds a new facet to the implications of the degradation of those compounds by Streptomyces, where linearization of surfactin was previously reported as a detoxification mechanism. Additionally, we hypothesize that lipopeptide-producing rhizobacteria and their biocontrol potential are impacted by the degradation of their lipopeptides as observed with the polarized motility of B. velezensis, avoiding the confrontation zone with Streptomyces and the loss of antifungal properties of degraded iturin. This work illustrates how CLPs, once released in the environment, may rapidly be remodeled or degraded by members of the bacterial community, with potential impacts on CLP-producing rhizobacteria and the biocontrol products derived from them.

microbiology↗

bioRxiv2023

Common scab disease: structural basis of elicitor recognition in pathogenic Streptomyces species

In Streptomyces scabiei, the main causative agent of common scab disease of root and tuber crops, the interaction between the substrate-binding protein (SBP) CebE (CebEscab) and cellotriose released by the plant host (KD in the nanomolar range) is the first event for the onset of its pathogenic lifestyle. Here we report the structure of CebEscab in complex with cellotriose at a 1.55 [A] resolution, adopting a general fold of the B subcluster of SBPs. The interaction between CebEscab and cellotriose involves multiple direct or water-mediated hydrogen bonds and hydrophobic interactions, the glucose monomer at the non-reducing end occupying the most conserved part of the substrate-binding cleft. As main interactions between the two domains of CebE involve cellotriose itself, the closed conformational state of CebE is performed via an induced-fit ligand binding mechanism where cellotriose binding triggers the domain movement. Analysis of regulon predictions revealed that the signaling pathway from the CebE-mediated cellotriose transport to the transcriptional activation of thaxtomin phytotoxin biosynthesis is conserved in Streptomyces spp causing common scab, except for Streptomyces ipomoeae that specifically colonizes sweet potatoes and responds to other and yet unknown virulence elicitors. Interestingly, strains belonging to pathogenic species turgidiscabies and caniscabies have a cellotriose-binding protein orthologous to the CebE protein of the saprophytic species Streptomyces reticuli with lower affinity for its substrate (KD in the micromolar range), suggesting higher cellotriose concentrations for perception of their host. Our work also provides the structural basis for the uptake of cellobiose and cellotriose by non-pathogenic cellulose-decomposing Streptomyces species. ImportanceCommon scab is a disease caused by few Streptomyces species that affects important root and tuber crops including potato, beet, radish, and parsnip, resulting in major economic losses worldwide. In this work we unveiled the molecular basis of host recognition by these pathogens by solving the structure of the sugar-binding protein CebE of S. scabiei in complex with cellotriose, the main elicitor of the pathogenic lifestyle of these bacteria. We further revealed that the signaling pathway from CebE-mediated transport of cellotriose is conserved in all pathogenic species except S. ipomoeae that causes soft rot disease on sweet potatoes. Our work also provides the structural basis of the uptake of cellobiose and cellotriose in saprophytic Streptomyces species, the first step activating the expression of the enzymatic system degrading the most abundant polysaccharide on earth, cellulose. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=125 SRC="FIGDIR/small/540135v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@1915dd1org.highwire.dtl.DTLVardef@a65aaforg.highwire.dtl.DTLVardef@18bc293org.highwire.dtl.DTLVardef@937d2f_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LICellotriose uptake triggers common scab in tuber/root crops by Streptomyces scabiei C_LIO_LICrystal structure of CebE of S. scabiei interacting with cellotriose is solved C_LIO_LICellotriose triggers the closed conformational state of CebE C_LIO_LIThe CebE/cellotriose route to pathogenicity is conserved in Streptomyces species C_LIO_LICebE-type background may affect the cellotriose concentration eliciting virulence C_LI

microbiology↗

bioRxiv2022

Lunaemycins, new cyclic hexapeptide antibiotics from the cave moonmilk-dweller Streptomyces lunaelactis MM109

Streptomyces lunaelactis strains have been isolated from moonmilk deposits which are calcium carbonate speleothems used for centuries in traditional medicine for their antimicrobial properties. Genome mining revealed that these strains are a remarkable example of a Streptomyces species with huge heterogeneity regarding their content in biosynthetic gene clusters (BGCs) for specialized metabolite production. BGC 28a is one of the cryptic BGCs that is only carried by a subgroup of S. lunaelactis strains for which in silico analysis predicted the production of nonribo-somal peptide antibiotics containing the non-proteogenic amino acid piperazic acid (Piz). Comparative metabolomics of culture extracts of S. lunaelactis strains either or not holding BGC 28a combined with MS/MS-guided peptidogenomics and 1H/13C NMR allowed to identify the cyclic hexapeptide with the amino acid sequence (D-Phe)-(L-HO-Ile)-(D-Piz)-(L-Piz)-(D-Piz)-(L-Piz), called lunaemycin A, as the main compound synthesized by BGC 28a. Molecular networking further identified 18 additional lunaemycins, 14 of them having their structure elucidated by HRMS/MS. Antimicrobial assays demonstrated a huge bactericidal activity of lunaemycins against Gram-positive bacteria including multi-drug resistant clinical isolates. Our work demonstrates how accurate in silico analysis of a cryptic BGC can highly facilitate the identification, the structural elucidation, and the bioactivity of its associated specialized metabolites.

microbiology↗

bioRxiv2022

AURTHO: autoregulation as facilitator of cis-acting element discovery of orthologous transcription factors

Transcriptional regulation is key in bacteria for providing an adequate response in time and space to changing environmental conditions. However, despite decades of research, the binding sites and therefore the target genes and the function of most transcription factors (TFs) remain unknown. Filling this gap in knowledge through conventional methods represents a colossal task which we demonstrate here can be significantly facilitated by a widespread feature in transcriptional control: the autoregulation of TFs implying that the yet unknown transcription factor binding site (TFBS) is neighbouring the TF itself. In this work, we describe the "AURTHO" methodology (AUtoregulation of oRTHOlogous transcription factors), consisting of analyzing upstream regions of orthologous TFs in order to uncover their associated TFBSs. AURTHO enabled the de novo identification of novel TFBSs with an unprecedented improvement in terms of quantity and reliability. DNA-protein interaction studies on a selection of candidate cis-acting elements yielded an >90% success rate, demonstrating the efficacy of AURTHO at highlighting true TF-TFBS couples and confirming the identification in a near future of a plethora of TFBSs across all bacterial species. Key pointsO_LITranscription factor (TF) autoregulation implies that their binding site (TFBS) is in their close vicinity C_LIO_LIWe developed and assessed the AURTHO methodology (AUtoregulation of oRTHOlogous TFs) for TFBS discovery C_LIO_LIOur results shows that AURTHO greatly facilitates the identification of highly reliable novel TFBSs C_LI

bioinformatics↗

bioRxiv2022

Structure and function of BcpE2, the most promiscuous GH3-family beta-glucosidase for scavenging glucose from heterosides

Cellulose being the most abundant polysaccharide on earth, beta-glucosidases hydrolyzing cello-oligosaccharides are key enzymes to fuel glycolysis in microorganisms developing on plant material. In Streptomyces scabiei, the causative agent of common scab in root and tuber crops, a genetic compensation phenomenon safeguards the loss of the gene encoding the cello-oligosaccharide hydrolase BglC by awakening the expression of alternative beta-glucosidases. Here we reveal that the BglC compensating enzyme BcpE2 is the GH3-family beta-glucosidase that displays the highest reported substrate promiscuity able to release the glucose moiety of all tested types of plant-derived heterosides (aryl {beta}-glucosides, monolignol glucosides, cyanogenic glucosides, anthocyanosides, and coumarin heterosides). BcpE2 structure analysis highlighted a large cavity in the PA14 domain that covers the active site, and the high flexibility of this domain would allow proper adjustment of this cavity for disparate heterosides. The exceptional substrate promiscuity of BcpE2 provides microorganisms a versatile tool for scavenging glucose from plant-derived nutrients that widely vary in size and structure. Importantly, scopolin is the only substrate commonly hydrolyzed by both BglC and BcpE2 thereby generating the potent virulence inhibitor scopoletin. Next to fueling glycolysis, both enzymes thus also interfere with the plant defense mechanisms to fine-tune the strength of virulence.

biochemistry↗

bioRxiv2021

BglC of Streptomyces scabiei is a scopolin-hydrolyzing β-glucosidase interfering with the host defense mechanism

The beta-glucosidase BglC fulfills multiple functions in both primary metabolism and induction of pathogenicity of Streptomyces scabiei, the causative agent of the common scab disease of root and tuber crops. Indeed, this enzyme hydrolyzes cellobiose and cellotriose to directly feed glycolysis with glucose, but also modifies the intracellular concentration of these cello-oligosaccharides which are the virulence elicitors. The inactivation of bglC also led to unexpected phenotypes such as the constitutive overproduction of thaxtomin A, the main virulence determinant of S. scabiei. In this work we revealed a new target substrate of BglC, the phytoalexin scopolin. Removal of the glucose moiety of scopolin generates scopoletin, a potent inhibitor of thaxtomin A production. The hydrolysis of scopolin by BglC presents substrate inhibition kinetics which contrasts with the typical Michaelis-Menten saturation curve previously observed for the degradation of its natural substrate cellobiose. Our work therefore reveals that BglC targets both cello-oligosaccharide elicitors emanating from the hosts of S. scabiei, and the scopolin phytoalexin generated by the host defense mechanisms, thereby occupying a key position to fine-tune the production of the main virulence determinant thaxtomin A.

microbiology↗

bioRxiv2021

The virulome of Streptomyces scabiei in response to cello-oligosaccharides elicitors

2.The development of spots or lesions symptomatic of the common scab disease on root and tuber crops is caused by few pathogenic Streptomyces with Streptomyces scabiei 87-22 as the model species. Thaxtomin phytotoxins are the primary virulence determinants, mainly acting by impairing cellulose synthesis, and their production in S. scabiei is in turn boosted by the cello-oligosaccharides released from host plants. In this work we aimed to determine which molecules and which biosynthetic gene clusters (BGCs) of the specialized metabolism of S. scabiei 87-22 show a production and/or transcriptional response to cello-oligosaccharides. Comparative metabolomic and transcriptomic analyses revealed that molecules of the virulome of S. scabiei induced by cellobiose and cellotriose include i) thaxtomins and concanamycins phytotoxins (and to a lesser extent N-coronafacoyl-L-isoleucine), ii) desferrioxamines, scabichelin and turgichelin siderophores in order to acquire iron essential for housekeeping functions, iii) ectoine for protection against osmotic shock once inside the host, and iv) bottromycins and concanamycins antimicrobials possibly to prevent other microorganisms from colonizing the same niche. Importantly, both cell-oligosaccharides reduced the production of the spore germination inhibitors germicidins and the plant growth regulators rotihibins. The metabolomic study also revealed that cellotriose is in general a more potent elicitor of the virulome compared to cellobiose. This result supports an earlier hypothesis that suggested that the trisaccharide would be the real virulence-triggering factor released from the plant cell wall through the action of thaxtomins. Interestingly, except for thaxtomins, none of these BGCs expression seems to be under direct control of the cellulose utilization repressor CebR suggesting the existence of another master regulator sensing the internalization of cello-oligosaccharides. Finally, we found nine additional cryptic and orphan BGCs that have their expression awakened by cello-oligosaccharides, demonstrating that other and yet to be discovered metabolites are part of the virulome of S. scabiei. 3. Impact statementUnveiling the environmental triggers that signal proper conditions for host colonization and what is the composition of the arsenal of metabolites specialized for this task (the virulome) is key to understand host-pathogen interactions. In this work, focused on the induction of the common scab disease caused by Streptomyces species, we provided further knowledge to both aspects i.e., i) highlighting the capability of cellotriose to trigger the entire virulome and not only the production of thaxtomin phytotoxins, and ii) identifying the set of metabolites that specifically respond to cello-oligosaccharides emanating from the plant under attack. Importantly, we also revealed that the expression of nine cryptic/orphan biosynthetic gene clusters (BGCs) involved in the production of unknown compounds was drastically activated upon cello-oligosaccharides import suggesting that a significant part of the virulome of S. scabiei remains to be discovered. Finally, we unexpectedly found that the expression control of most of the known and cryptic BGCs does not depend on the cello-oligosaccharide utilization repressor CebR which suggests the existence of another and yet unknown master regulator of the virulence in S. scabiei. 4. Significance as a BioResource to the communityNot Applicable 5. OutcomeNot Applicable 6. Data summary[A section describing all supporting external data including the DOI(s) and/or accession numbers(s), and the associated URL.] The authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files. RNAseq data were publicly deposited, and our experimental and analytical pipeline were described on the GEO database repository (Accession number: GSE181490)

microbiology↗

bioRxiv2019

Bagremycin Antibiotics and Ferroverdin iron-chelators are synthetized by the Same Gene Cluster

Biosynthetic gene clusters (BGCs) are organized groups of genes involved in the production of specialized metabolites. Typically, one BGC is responsible for the production of one or several similar compounds with bioactivities that usually only vary in terms of strength and/or specificity. Here we show that the previously described ferroverdins and bagremycins, which are families of metabolites with different bioactivities, are produced from the same BGC, whereby the fate of the biosynthetic pathway depends on iron availability. Under conditions of iron depletion, the monomeric bagremycins are formed, which are amino-aromatic antibiotics resulting from the condensation of 3-amino-4-hydroxybenzoic acid with p-vinylphenol. Conversely, when iron is abundantly available, the biosynthetic pathway additionally produces a molecule based on p-vinylphenyl-3-nitroso-4-hydroxybenzoate, which complexes iron to form the trimeric ferroverdins that have anticholesterol activity. Thus our work challenges the concept that BGCs should produce a single family of molecules with one type of bioactivity, the occurrence of the different metabolites being triggered by the environmental conditions.

microbiology↗

bioRxiv2018

Prodiginine Production in Streptomyces coelicolor Correlates Temporally and Spatially to Programmed Cell Death

Programmed cell death (PCD) is a common feature of multicellularity and morphogenesis in bacteria. While cell death has been well documented when Streptomyces species switch from vegetative (nutrition) to aerial (reproduction) growth, lethal determinants are yet to be discovered to unveil the genetic basis of PCD in mycelial bacteria. In this work we used prodiginines of Streptomyces coelicolor as model to test the hypothesis that a bacterium uses self-made antiproliferative DNA-damaging agents as toxins of their PCD process. Spatio-temporal visualisation of the autofluorescence of prodiginines reveals that their biosynthesis is triggered in the dying zone of the colony prior to morphological differentiation of the mycelium. A prodiginine nonproducer showed hyper-accumulation of viable filaments, with increased RNA and proteins synthesis when most of the mycelium of the wild-type strain was dead when prodiginine accumulated. Addition of a prodiginine synthesis inhibitor also strongly favoured viable over dead filaments. As self-toxicity has also been reported for other producers of DNA-damaging agents we propose that cytotoxic metabolites synthetized during the morphological transition of filamentous bacteria may be used to execute PCD.\n\nSignificance StatementActinobacteria are prolific producers of compounds with antiproliferative activity, but why these bacteria synthetize metabolites with this bioactivity has so far remained a mystery. Using prodiginines (PdGs) as model system, we revealed that the spatio-temporal synthesis of these molecules correlates to cell death of the producer Streptomyces coelicolor and that inhibition of their synthesis results in hyper-accumulation of viable filaments. Since PdGs potentiate death of S. coelicolor recurrently prior to morphological differentiation, this is a form of programmed cell death (PCD). Hence, next to weapons in competition between organisms or signals in inter- and intra-species communications, we propose a third role for secondary metabolites i.e., elements required for self-toxicity in PCD processes.

microbiology↗

bioRxiv2017

Contribution of the β-glucosidase BglC to the Onset of the Pathogenic Lifestyle of Streptomyces scabies

Common scab disease on root and tuber plants is caused by Streptomyces scabies and related species which use the cellulose synthase inhibitor thaxtomin A as main phytotoxin. Thaxtomin production is primarily triggered by the import of cello-oligosaccharides. Once inside the cell, the fate of the cello-oligosaccharides is dichotomized into i) fueling glycolysis with glucose for the saprophytic lifestyle through the action of {beta}-glucosidase(s) (BG), and ii) eliciting the pathogenic lifestyle by inhibiting the CebR-mediated transcriptional repression of thaxtomin biosynthetic genes. Here we investigated the role of scab57721 encoding a putative BG (BglC) in the onset of the pathogenicity of S. scabies. Enzymatic assays showed that BglC was able to release glucose from cellobiose, cellotriose and all other cello-oligosaccharides tested. Its inactivation resulted in a phenotype opposite to what was expected as we monitored reduced production of thaxtomin when the mutant was cultivated on media containing cello-oligosaccharides as unique carbon source. This unexpected phenotype could be attributed to the highly increased activity of alternative intracellular BGs, probably as a compensation of bglC inactivation, which then prevented cellobiose and cellotriose accumulation to reduce the activity of CebR. In contrast, when the bglC null mutant was cultivated on media devoid of cello-oligosaccharides it instead constitutively produced thaxtomin. This observed hypervirulent phenotype does not fit with the proposed model of the cello-oligosaccharide-mediated induction of thaxtomin production and suggests that the role of BglC in the route to the pathogenic lifestyle of S. scabies is more complex than currently presented.

microbiology↗

bioRxiv2017

Qinichelins, novel catecholate-hydroxamate siderophores synthesized via a multiplexed convergent biosynthesis pathway

The explosive increase in genome sequencing and the advances in bioinformatic tools have revolutionized the rationale for natural product discovery from actinomycetes. In particular, this has revealed that actinomycete genomes contain numerous orphan gene clusters that have the potential to specify many yet unknown bioactive specialized metabolites, representing a huge unexploited pool of chemical diversity. Here, we describe the discovery of a novel group of catecholate-hydroxamate siderophores termed qinichelins (2-5) from Streptomyces sp. MBT76. Correlation between the metabolite levels and the protein expression profiles identified the biosynthetic gene cluster (BGC; named qch) most likely responsible for qinichelin biosynthesis. The structure of the molecules was elucidated by bioinformatics, mass spectrometry and NMR. Synthesis of the qinichelins requires the interplay between four gene clusters, for its synthesis and for precursor supply. This biosynthetic complexity provides new insights into the challenges scientists face when applying synthetic biology approaches for natural product discovery.\n\nPride repository reviewer account details:\n\nURL: https://www.ebi.ac.uk/pride/archive/login\n\nProject accession: PXD006577\n\nUsername: reviewer35793@ebi.ac.uk\n\nPassword: 3H0iM1FK

microbiology↗