03/01/2021
Systematic Discovery of Pseudomonad Genetic Factors Involved in Sensitivity to Tailocins
Gene content, however, cannot completely explain sensitivity.
The Science
Researchers in the Ecosystems and Networks Integrated with Genes and Molecular Assemblies (ENIGMA) project are investigating tailocin (phage tail–like bacteriocins) sensitivity factors. The research team aims to identify mechanisms behind resistance to self-intoxication.
The Impact
The researchers are seeking to reach conclusions by using resources previously developed by them and their collaborators. They are iidentifying tailocin gene clusters in Pseudomonas genomes and characterizing induced particles; using receptor (RB)-TnSeq libraries to identify mutants with altered fitness in the presence of tailocins; specifically assessing O-specific antigen (OSA) composition as well as phospholipid transport; and profiling tailocin sensitivity in 130 sequenced Pseudomonas isolates in reference to OSA composition.
Summary
The ENIGMA project is the first systematic effort to identify genetic factors involved in sensitivity to tailocins. The team has identified genes related to tailocin infectivity in the lipopolysaccharide (LPS) core and OSA biosynthetic gene clusters. They have found genes encoding outer membrane lipid asymmetry and LPS transport that are involved in sensitivity to tailocins. Strains with the same overall OSA cluster typically been found to display the same tailocin sensitivity pattern, but the researchers believe that gene content cannot completely explain sensitivity.
Principal Investigator(s)
Paul Adams
Lawrence Berkeley National Laboratory
[email protected]
Funding
The authors acknowledge the work of ENIGMA Scientific Focus Area (SFA) colleagues on isolating and sequencing the Pseudomonas spp. reported here, specifically: A. Aaring, E. J. Alm, M. Callaghan, H. K. Carlson, J. V. Kuehl, R. A. Melnyk, and S. J. Spencer. The U.S. Department of Energy (DOE) Office of Science acknowledges B. A. Adler for discussions and critical feedback on the manuscript. This work was funded by ENIGMA, an SFA Program at Lawrence Berkeley National Laboratory (LBNL), supported by the Office of Biological and Environmental Research (BER), within the DOE Office of Science, under contract DE-AC02-05CH11231. Proteomics data acquisition and analysis was performed at the Vincent J. Proteomics/Mass Spectrometry Laboratory at the University of California(UC)–Berkeley (UCB), supported in part by the National Institutes of Health (NIH) S10 Instrumentation Grant S10RR025622. PacBio genome sequence and BarSeq data acquisition were performed at the Vincent J. Coates Genomics Sequencing Laboratory at the UCB, supported by NIH S19 Instrumentation Grants S10RR029668, S10RR027303, and OD018174. The DOE Office of Science acknowledges support from the U.S. National Science Foundation (NSF) Graduate Research Fellowship Program under Grant No. DGE 1752814, the Croucher Foundation through a Croucher Scholarship for Doctoral Study, and the Drs. Richard Charles and Esther Yewpick Lee Charitable Foundation through an R C Lee Centenary Scholarship. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of NSF.
References
Carim, S. et al. “Systematic Discovery of Pseudomonad Genetic Factors Involved in Sensitivity to Tailocins.” The ISME Journal 15, 2289–2305 (2021). [DOI:10.1038/s41396-021-00921-1]