Charlton, S. G. V. | bioRxiv Science

bioRxiv2024

Single-cell approach dissecting agr quorum sensing dynamics in Staphylococcus aureus

Staphylococcus aureus both colonizes humans and causes severe virulent infections. Virulence is regulated by the agr quorum sensing system and its autoinducing peptide (AIP), with dynamics at the single-cell level across four agr-types - each defined by distinct AIP sequences and capable of cross-inhibition - remaining elusive. Employing microfluidics, time-lapse microscopy, and deep-learning image analysis, we uncovered significant differences in AIP sensitivity among agr-types. We observed bimodal agr activation, attributed to intergenerational phenotypic stability and influenced by AIP concentration. Upon AIP stimulation, agr-III showed AIP insensitivity, while agr-II exhibited increased sensitivity and prolonged generation time. Beyond expected cross-inhibition of agr-I by heterologous AIP-II and -III, the presumably cross-activating AIP-IV also inhibited agr-I. Community interactions across different agr-type pairings revealed four main patterns: stable or switched dominance, and delayed or stable dual activation, influenced by community characteristics. These insights underscore the potential of personalized treatment strategies considering virulence and genetic diversity.

microbiology↗

bioRxiv2023

Tunable living bacterial networks: A multi-scaled description of their self-assembly and mechanical behaviour

Bacterial aggregate formation and surface accumulation are increasingly viewed as alternative pathways for biofilm colonization. However, little is known about the dynamics of bacterial aggregate cluster-cluster assembly and their subsequent microstructural and mechanical properties. To this end, we studied experimentally and computationally an aggregating bacterial system that forms a space-spanning interconnected network via cluster-cluster assembly. By controllably inducing bacterial filamentation, we aimed to understand how cell length distribution and cell surface hydrophobicity control the dynamics of aggregation and sedimentation, as well as the microstructure and mechanics of the settled bacterial networks. We found that filamentation lowers the percolation threshold, leading to gelation at a lower number density with distinct assembly dynamics and lower network connectivity. Furthermore, we analyzed the mechanical properties of the bacterial networks. Static stress tests reveal three yielding modes: discrete cluster-cluster disassembly, collective delamination, and sub-regional network fracture. The yielding modes are consistent with the gel-like viscoelastic properties of the cluster-cluster assembled networks observed during macroscale rheometry. In particular, we observe a scaling relationship between the storage modulus and the volume fraction, characteristic of an attractive rod gel. Our experimental observations are supported by Langevin dynamic simulations, providing mechanistic insights into the factors determining network self-assembly and connectivity. Our findings elucidate the gel-like structure-function dynamics in cluster-cluster aggregated bacterial systems and underscore the fundamental importance of filamentation in their properties and mechanical behavior.

biophysics↗