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Gueguen-Chaignon, V.

Publications and source records attributed to Gueguen-Chaignon, V..

4 recordsLinked to original sources

The functional plasticity of the YhdWXYZ ABC transporter enables antibiotic homeostasis and host colonisation in enteric bacteria

ABC transporters are key determinants of bacterial adaptation, yet their functional plasticity remains poorly understood. Here, we characterize the type I ABC transporter YhdWXYZ and uncover a striking functional divergence linked to the presence of its substrate-binding protein (SBP). In Escherichia coli K-12, where yhdW is a pseudogene, deletion of yhdWXYZ increases susceptibility to mecillinam and lomefloxacin and leads to intracellular accumulation of lomefloxacin, indicating a role in antibiotic homeostasis. In contrast, in Citrobacter rodentium, which encodes a complete YhdWXYZ system, deletion of the transporter does not affect antibiotic susceptibility. Biochemical analyses demonstrate that the YhdW SBP of C. rodentium binds asparagine with high affinity; however, genetic and physiological assays indicate that YhdWXYZ is not a primary asparagine importer under laboratory conditions, suggesting redundancy with other transport systems. Importantly, in vivo infection experiments reveal that YhdWXYZ contributes to early colonization and persistence in the host, as mutants display reduced bacterial loads and altered intestinal pathology in mice. Together, these findings show that loss of the SBP in E. coli is associated with a functional shift of YhdWXYZ toward antibiotic homeostasis, whereas in C. rodentium, the complete transporter contributes to host adaptation. This work highlights the evolutionary and functional flexibility of ABC transporters in bacterial physiology and pathogenesis.

microbiology↗

A DltE-DltD-DltX interaction network regulates lipoteichoic acid D-alanylation in Lactiplantibacillus plantarum and symbiotic drosophila growth promotion

D-alanylation of teichoic acids is a widespread modification of Gram-positive bacterial cell envelopes that modulates resistance to environmental stresses and host interactions. Although the cytosolic steps of this pathway are well characterized, the extracellular reactions responsible for transferring D-alanine onto teichoic acids remain poorly understood. Here we investigate the role of DltD in the commensal bacterium Lactiplantibacillus plantarum. We determined the 2.3 [A] crystal structure of the extracellular catalytic domain of DltD, which adopts an SGNH-hydrolase fold with a conserved Ser-His-Asp catalytic triad. Docking analyses with lipoteichoic acids (LTA) fragments suggest that the glycerol-phosphate backbone of LTA is accommodated along a surface groove leading to the catalytic serine, with conserved residues contributing to substrate positioning. Biochemical measurements further reveal direct interactions between DltD, the acyl-carrier protein DltX, and the LTA esterase DltE. The conserved C-terminal motif of DltX binds DltD and is required for efficient D-alanylation and for L. plantarum-mediated promotion of Drosophila juvenile growth. Together, these findings support a DltX-dependent acyl-transfer mechanism and reveal an interaction network that coordinates LTA D-alanylation in a symbiotic bacterium.

biochemistry↗

Structure-Function analysis of Lactiplantibacillus plantarum DltE reveals D-alanylated lipoteichoic acids as direct symbiotic cues supporting Drosophila juvenile growth

Metazoans establish mutually beneficial interactions with their resident microorganisms. However, our understanding of the microbial cues contributing to host physiology remains elusive. Previously, we identified a bacterial machinery encoded by the dlt operon involved in Drosophila melanogasters juvenile growth promotion by Lactiplantibacillus plantarum. Here, using crystallography combined with biochemical and cellular approaches, we investigate the physiological role of an uncharacterized protein (DltE) encoded by this operon. We show that LTAs but not WTAs are D-alanylated in Lactiplantibacillus plantarumNC8 cell envelope and demonstrate that DltE is a D-Ala carboxyesterase removing D-Ala from LTA. Using the mutualistic association of L. plantarumNC8 and Drosophila melanogaster as a symbiosis model, we establish that D-Ala-LTAs are direct symbiotic cues supporting intestinal peptidase expression and juvenile growth in Drosophila. Our results pave the way to probing the contribution of D-Ala-LTA to host physiology in other symbiotic models.

microbiology↗

The unique Brucella effectors NyxA and NyxB target SENP3 to modulate the subcellular localisation of nucleolar proteins

The cell nucleus is a primary target for intracellular bacterial pathogens to counteract immune responses and hijack host signalling pathways to cause disease. The mechanisms controlling nuclear protein localisation in the context of stress responses induced upon bacterial infection are still poorly understood. Here we show that the Brucella abortus effectors NyxA and NyxB interfere with the host sentrin specific protease 3 (SENP3), which is essential for intracellular replication. Translocated Nyx effectors directly interact with SENP3 via a defined acidic patch identified from the crystal structure of NyxB, preventing its nucleolar localisation at the late stages of the infection. By sequestering SENP3, the Nyx effectors induce the cytoplasmic accumulation of the nucleolar AAA-ATPase NVL, the large subunit ribosomal protein L5 (RPL5) and the ribophagy receptor NUFIP1 in Nyx-enriched structures in the vicinity of replicating bacteria. This shuttling of ribosomal biogenesis-associated nucleolar proteins is negatively regulated by SENP3 and dependent on the autophagy-initiation protein Beclin1, indicative of a ribophagy-derived process induced during Brucella infection. Our results highlight a new nucleomodulatory function by two unique Brucella effectors, and reveal that SENP3 is a critical regulator of the subcellular localisation of multiple nucleolar proteins during Brucella infection, promoting intracellular replication.

microbiology↗