Acquired Type III Secretion System Determines Environmental Fitness of Epidemic Vibrio parahaemolyticus in the Interaction with Bacterivorous Protists

Genome analyses of marine microbial communities have revealed the widespread occurrence of genomic islands (GIs), many of which encode for protein secretion machineries described in the context of bacteria-eukaryote interactions. Yet experimental support for the specific roles of such GIs in aquatic community interactions remains scarce. Here, we test for the contribution of type III secretion systems (T3SS) to the environmental fitness of epidemic Vibrio parahaemolyticus. Comparisons of V. parahaemolyticus wild types and T3SS-defective mutants demonstrate that the T3SS encoded on genome island VPaI-7 (T3SS-2) promotes survival of V. parahaemolyticus in the interaction with diverse protist taxa. Enhanced persistence was found to be due to T3SS-2 mediated cytotoxicity and facultative parasitism of V. parahaemolyticus on coexisting protists. Growth in the presence of bacterivorous protists and the T3SS-2 genotype showed a strong correlation across environmental and clinical isolates of V. parahaemolyticus. Short-term microcosm experiments provide evidence that protistan hosts facilitate the invasion of T3SS-2 positive V. parahaemolyticus into a coastal plankton community, and that water temperature and productivity further promote enhanced survival of T3SS-2 positive V. parahaemolyticus. This study is the first to describe the fitness advantage of GI-encoded functions in a microbial food web, which may provide a mechanistic explanation for the global spread and the seasonal dynamics of V. parahaemolyticus pathotypes, including the pandemic serotype cluster O3:K6, in aquatic environments

Identification of Motifs of <em>Burkholderia pseudomallei</em> BimA Required for Intracellular Motility, Actin Binding, and Actin Polymerization

Actin-based motility of the melioidosis pathogen Burkholderia pseudomallei requires BimA (Burkholderia intracellular motility A). The mechanism by which BimA mediates actin assembly at the bacterial pole is ill-defined. Toward an understanding of the regions of B. pseudomallei BimA required for intracellular motility and the binding and polymerization of actin, we constructed plasmid-borne bimA variants and glutathione-S-transferase fusion proteins with in-frame deletions of specific motifs. A 13-amino-acid direct repeat and IP(7) proline-rich motif were dispensable for actin binding and assembly in vitro, and expression of the mutated proteins in a B. pseudomallei bimA mutant restored actin-based motility in J774.2 murine macrophage-like cells. However, two WASP homology 2 (WH2) domains were found to be required for actin binding, actin assembly, and plaque formation. A tract of five PDASX direct repeats influenced the polymerization of pyrene-actin monomers in vitro and was required for actin-based motility and intercellular spread, but not actin binding. None of the mutations impaired surface expression or polar targeting of BimA. The number of PDASX repeats varied in natural isolates from two to seven. Such repeats acted additively to promote pyrene-actin polymerization in vitro, with stepwise increases in the rate of polymerization as the number of repeats was increased. No differences in the efficiency of actin tail formation could be discerned between strains expressing BimA variants with two, five, or seven PDASX repeats. The data provide valuable new insights into the role of conserved and variable motifs of BimA in actin-based motility and intercellular spread of B. pseudomallei

Safe and just Earth system boundaries.

This is the final version. Available from Nature Research via the DOI in this record. Data availability The data supporting Figs. 2 and 3 are available at https://doi.org/10.6084/m9.figshare.22047263.v2 and https://doi.org/10.6084/m9.figshare.20079200.v2, respectively. We rely on other published datasets for the climate boundary16, N boundary72 (model files are at https://doi.org/10.5281/zenodo.6395016), phosphorus73,74 (scenario breakdowns are at https://ora.ox.ac.uk/objects/uuid:d9676f6b-abba-48fd-8d94-cc8c0dc546a2, and a summary of agricultural sustainability indicators is at https://doi.org/10.5281/zenodo.5234594), current N surpluses129,130 (the repository at https://dataportaal.pbl.nl/downloads/IMAGE/GNM) with the critical N surplus limit72 subtracted, and estimated subglobal P concentration in runoff based on estimated P load to freshwater131 and local runoff data132,133. Current functional integrity is calculated from the European Space Agency WorldCover 10-metre-resolution land cover map (https://esa-worldcover.org/en). The safe boundary and current state for groundwater are derived from the Gravity Recovery And Climate Experiment (http://www2.csr.utexas.edu/grace/RL06_mascons.html) and the Global Land Data Assimilation System (https://disc.gsfc.nasa.gov/datacollection/GLDAS_NOAH025_3H_2.1.html). More information is available in ‘Code availability’ and Supplementary Methods. Source data for Fig. 2 are provided with this paper.Code availability: The code used to produce Figs. 2 and 3 are available at https://doi.org/10.6084/m9.figshare.22047263.v2 and https://doi.org/10.6084/m9.figshare.20079200.v2, respectively. The code used to make the nutrient Earth system boundary layers in Fig. 3 is available at https://doi.org/10.5281/zenodo.7636716. The code used to make the surface water layer in Fig. 3 and derive the subglobal Earth system boundaries for surface water is available at https://doi.org/10.5281/zenodo.7674802. The code to estimate current functional integrity is available at https://figshare.com/articles/software/integrity_analysis/22232749/2. The code to derive the groundwater layer in Fig. 3 and derive the total annual groundwater recharge is available at https://doi.org/10.5281/zenodo.7710540.The stability and resilience of the Earth system and human well-being are inseparably linked1-3, yet their interdependencies are generally under-recognized; consequently, they are often treated independently4,5. Here, we use modelling and literature assessment to quantify safe and just Earth system boundaries (ESBs) for climate, the biosphere, water and nutrient cycles, and aerosols at global and subglobal scales. We propose ESBs for maintaining the resilience and stability of the Earth system (safe ESBs) and minimizing exposure to significant harm to humans from Earth system change (a necessary but not sufficient condition for justice)4. The stricter of the safe or just boundaries sets the integrated safe and just ESB. Our findings show that justice considerations constrain the integrated ESBs more than safety considerations for climate and atmospheric aerosol loading. Seven of eight globally quantified safe and just ESBs and at least two regional safe and just ESBs in over half of global land area are already exceeded. We propose that our assessment provides a quantitative foundation for safeguarding the global commons for all people now and into the future.Stockholm Universit