Search bioRxiv⌕ Search

Biology subjects

Borde, C.

Publications and source records attributed to Borde, C..

2 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↗

DNA topoisomerase I acts as supercoiling sensor for transcription elongation in E. coli

When DNA is transcribed to RNA, the DNA double helix is constantly unwound and rewound to provide access for RNA polymerase (RNAP). This induces DNA supercoiling as a function of transcript length due to over- and under-twisting of the DNA downstream and upstream of RNAP, respectively. Using single-particle cryo-EM and in vivo assays we investigated the relationship between bacterial RNAP and DNA Topoisomerase I (TopoI), which removes negative supercoils accumulating upstream of RNAP. TopoI binds to relaxed DNA upstream of RNAP in a manner suggesting a sensory role awaiting the formation of negative supercoils and involving a conformational switch in the functional domains of TopoI. On DNA substrates mimicking negatively supercoiled DNA, TopoI threads one strand into the active site for cleavage while binding the complementary strand with an auxiliary domain. We propose a comprehensive model for DNA relaxation in the context of a transcribing RNAP.

molecular biology↗