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Brown, L. M.

Publications and source records attributed to Brown, L. M..

2 recordsLinked to original sources

Pseudomonas aeruginosa PA14 biofilms produce R-bodies, extendable protein polymers with roles in host colonization and virulence

Pseudomonas aeruginosa PA14, an opportunistic pathogen of diverse hosts, contains genes with the potential to confer production of R-bodies (i.e., a "reb cluster"). R-bodies are large, extendable protein polymers best known for their role in killing of paramecia by the bacterium Caedibacter taeniospiralis, and genes in the reb cluster have been implicated in PA14 virulence. Here, we present evidence that PA14 expresses reb cluster genes during colonization of plant and nematode hosts. We identify products of the reb cluster that are R-body-associated and that control stochastic expression of R-body structural genes. We also show that R-body production is required for full virulence in nematodes. Analyses of nematode ribosome content and immune response indicate that R-bodies act via a mechanism involving ribosome cleavage and translational inhibition. These observations provide insight into the biology of R-body production and its consequences during P. aeruginosa infection.

microbiology

Formation of biomolecular condensates in bacteria by tuning protein electrostatics

Biomolecular condensates provide a strategy for cellular organization without a physical membrane barrier while allowing for dynamic, responsive organization of the cell. To date, very few biomolecular condensates have been identified in prokaryotes, presenting an obstacle to engineering these compartments in bacteria. As a novel strategy for bacterial compartmentalization, protein supercharging and complex coacervation were employed to engineer liquid-like condensates in E. coli. A simple model for the phase separation of supercharged proteins was developed and used to predict intracellular condensate formation. Herein, we demonstrate that GFP-dense condensates formed by expressing GFP variants of sufficient charge in cells are dynamic and enrich specific nucleic acid and protein components. This study provides a fundamental characterization of intracellular phase separation in E. coli driven by protein supercharging and highlights future utility in designing functional synthetic membraneless organelles.

synthetic biology