An intrinsic mechanism for coordinated production of the contact-dependent and contact-independent weapon systems in a soil bacterium

Abstract

Soil bacteria possess multiple weapons to fend off microbial competitors. Currently, we poorly understand the factors guiding bacterial decisions about weapon systems deployment. In this study, we investigated how such decisions are made by the soil bacterium Lysobacter enzymogenes, used in antifungal plant protection. We found that weapons production is guided by environmental cues. In rich media, which likely mimic environments crowded with other microbes, L. enzymogenes produces a contact-dependent weapon, type six secretion system (T6SS). In nutrient-poor media, likely dominated by filamentous oomycetes and fungi, L. enzymogenes synthesizes and secretes a heat-stable antifungal factor (HSAF), a contact-independent weapon. Surprisingly, the T6SS inner tube protein Hcp is accumulated intracellularly even in nutrient-poor media, when the T6SS is not assembled. We found that Hcp interacts with the transcription factor Clp required for activating HSAF biosynthesis operon expression. Hcp protects Clp from binding to c-di-GMP, an intracellular second messenger inhibiting DNA binding. The increased concentration of c-di-GMP-free Clp thus leads to higher gene expression and HSAF production. Therefore, when the contact-dependent weapon, T6SS, is not in use, accumulation of one of its structural components, Hcp, serves as a signal to enhance production of the contact-independent weapon, HSAF. The uncovered environment-dependent and auto-regulatory mechanisms shed light on the processes governing deployment of various weapon systems in environmental bacteria.

Author summary

Soil bacteria face competition from diverse microbial species. To stay competitive, they deploy a variety of weapons. At present, we know little about factors influencing decisions about which weapons to produce at any given time, and about mechanisms through which these decisions are carried out. In this study, we show that in the soil bacterium, Lysobacter enzymogenes, synthesis of the contact-dependent weapon, known as type six secretion system (T6SS) occurs under different conditions, compared to those conductive to the production of the contact-independent weapon, toxin HSAF. Further, when T6SS is not assembled, one of its structural components, Hcp, coactivates HSAF operon expression and HSAF synthesis. This study reveals that decisions about contact-dependent and contact-independent weapon production in bacteria are governed by both environmental cues and intrinsic coordination mechanisms.