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Biology subjects

Berruto, C.

Publications and source records attributed to Berruto, C..

2 recordsLinked to original sources

A 3D-Printed Capillary Tube Holder for High-Throughput Chemotaxis Assays

Bacterial chemotaxis is an important behavior to study to understand spatial segregation of species in mixed communities and the assembly of host microbiomes. This is particularly relevant in the rhizosphere, where chemoattraction towards root exudates is an important determinate of plant colonization. However, current methods to screen chemoeffectors are limited in their throughput, creating a barrier to generating comprehensive datasets describing chemotactic profiles for species of interest. Here, we describe a novel, 3D-printed capillary tube holder approach, which enables up to 384 simultaneous capillary tube chemotaxis assays. We optimized and benchmarked our assay using Escherichia coli k12 and Bacillus subtilis 3610 and their known chemoattractants: serine and aspartate. We then tested the threshold concentration of these chemoattractants in our assay and found we could detect chemoattraction towards concentrations spanning multiple orders of magnitudes. In this paper, we describe our novel high-throughput chemotaxis assay in detail and provide the necessary files for 3D printing the capillary tube holder.

microbiology↗

Targeted protein evolution in the gut microbiome by diversity-generating retroelements

Diversity-generating retroelements (DGRs) accelerate evolution by rapidly diversifying variable proteins. The human gastrointestinal microbiota harbors the greatest density of DGRs known in nature, suggesting they play adaptive roles in this environment. We identified >1,100 unique DGRs among human-associated Bacteroides species and discovered a subset that diversify adhesive components of Type V pili and related proteins. We show that Bacteroides DGRs are horizontally transferred across species, that some are highly active while others are tightly controlled, and that they preferentially alter the functional characteristics of ligand-binding residues on adhesive organelles. Specific variable protein sequences are enriched when Bacteroides strains compete with other commensal bacteria in gnotobiotic mice. Analysis of >2,700 DGRs from diverse phyla in mother-infant pairs shows that Bacteroides DGRs are preferentially transferred to vaginally delivered infants where they actively diversify. Our observations provide a foundation for understanding the roles of stochastic, targeted genome plasticity in shaping host-associated microbial communities.

microbiology↗