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Desguin, B.

Publications and source records attributed to Desguin, B..

3 recordsLinked to original sources

Chemical dysbiosis byproducts trigger predation via alternative activation of a peptide quorum sensor in salivarius streptococci

Cell-to-cell communication in Gram-positive bacteria is predominantly orchestrated by cytoplasmic sensors of the RRNPPA family. To date, all characterized members of this family are activated by small, unmodified peptide pheromones that mediate bacterial signaling. In the human commensal Streptococcus salivarius, the RRNPPA sensor ComR controls both competence (DNA transformation) and predation (bacteriocin production). Here, we reveal that ComR can be dually activated by its cognate peptide (XIP) and a distinct class of small organic molecules. A targeted screen of [~]200 organic compounds identified hydroxyphenylacetic acid (HPAA), a bacterial dysbiosis byproduct accumulating in human fluids, as a potent inducer of ComR. Using in vivo and in vitro approaches, we demonstrated that HPAA and structurally related carboxylic acids derived from bulky hydrophobic amino acids bind the pheromone-accommodating pocket, leading to ComR activation. Strikingly, while XIP-mediated activation is transient and regulates both competence and predation, HPAA induces a sustained, predation-oriented response. Furthermore, we showed that Porphyromonas gingivalis, an oral pathogen, produces sufficient (H)PAA quantity to trigger bacteriocin production in S. salivarius, revealing a previously unrecognized chemical interplay between oral microbiota members. These findings highlight the remarkable versatility of cytoplasmic sensors to integrate diverse environmental cues, shedding new light on bacterial peptide-based communication and microbial homeostasis in the human microbiome. IMPORTANCEBacterial communication through quorum sensing (QS) is crucial for coordinating key physiological processes. While QS in Gram-positive bacteria has been predominantly associated with peptide pheromones, our study uncovers an undescribed alternative signaling mechanism. We demonstrate that ComR, a cytoplasmic receptor of the RRNPPA family, can be activated by its canonical peptide signal and by small organic molecules derived from the anaerobic breakdown of hydrophobic amino acids. This alternative activation pathway enhances the ability of S. salivarius to respond to microbial dysbiosis, maintaining ecological balance in the digestive tract. Beyond revealing a novel layer of bacterial communication, our findings suggest that many RRNPPA-family receptors previously considered "orphan" may respond to yet-undiscovered chemical signals. This work expands our understanding of bacterial sensing and opens new avenues for modulating microbial interactions in health and disease.

microbiology↗

Structures of two LarA-like nickel-pincer nucleotide cofactor-utilizing enzymes with a single catalytic histidine residue

The nickel pincer nucleotide (NPN) cofactor catalyzes the racemization/epimerization of -hydroxy acids in enzymes of the LarA family. The established proton-coupled hydride transfer mechanism requires two catalytic histidine residues that alternately act as general acids and general bases. Notably, however, a fraction of LarA homologs (LarAHs) lack one of the active site histidine residues, replacing it with an asparaginyl side chain that cannot participate in acid/base catalysis. Here, we investigated two such LarAHs and solved their cryo-electron microscopic structures with and without loaded NPN cofactor, respectively. The structures revealed a consistent octameric assembly that is unprecedented in the LarA family and unveiled a new set of active site residues that likely recognize and process substrates differently from those of the well-studied LarAHs. Genomic context analysis suggested their potential involvement in carbohydrate metabolism. Together, these findings lay the groundwork for expanding the breadth of reactions and the range of mechanisms of LarA enzymes.

biochemistry↗

Structural Basis for Catalysis and Substrate Specificity of a LarA Racemase with a Broad Substrate Spectrum

The LarA family consists of diverse racemases/epimerases that interconvert the diastereomers of a variety of -hydroxyacids by using a nickel-pincer nucleotide (NPN) cofactor. The hidden redox reaction catalyzed by the NPN cofactor makes LarA enzymes attractive engineering targets for applications. However, how a LarA enzyme binds its natural substrate and recognizes different -hydroxyacids has not been elucidated. Here, we report three high-resolution structures of the enzyme-substrate complexes of a broad-spectrum LarA enzyme from Isosphaera pallida (LarAIp). The substrate binding mode reveals an optimal orientation and distance between the hydride donor and acceptor, strongly supporting the proposed proton-coupled hydride transfer mechanism. The experimentally solved structures, together with the structural models of other LarA enzymes, allow us to identify the residues/structural elements critically involved in the interactions with different -hydroxyacid substrates. Collectively, this work provides a critical structural basis for catalysis and substrate recognition of the diverse enzymes in the LarA family, thus building a foundation for enzyme engineering.

biochemistry↗