Poster Presentation 50th Lorne Proteins Conference 2025

Genetic variation in individuals from a population of the minimalist bacteriophage Merri-merri-uth nyilam marra-natj driving evolution of the virus (#412)

Tze Young Thung 1 , Kher Shing Tan 1 , Alex Hall 1 , Murray White 2 , Rebecca Bamert 3 , Cara Press 1 , Francesca Short 1 , Rhys Dunstan 1 , Trevor Lithgow 1
  1. Department of Microbiology, Monash University, Clayton, VIC, Australia
  2. Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania , United States
  3. Infection & Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia

In a survey of a waterway on Wurundjeri land, two sub-populations of the bacteriophage Merri-merri-uth nyilam marra-natj (phage MMNM) were isolated on a permissive host, Klebsiella B5055 of capsule-type K2, but were distinguished by minor phenotypic differences. O-antigen side chains on the lipopolysaccharide layer of the outer membrane was identified as a receptor for phage binding. The variant phage MMNM(Ala134) showed an inhibited activity against Klebsiella AJ174-2, and this was used as a basis to select for further variation through experimental evolution. Over the course of an evolution experiment, 18 phages that evolved distinct phenotypes in terms of the morphologies of plaques formed when they infected host Klebsiella were subject to whole genome sequencing. The evolved phages had mutations in a small set of proteins that contribute to the baseplate portion of the phage virion. Phages MMNM and MMNM(Ala134) are minimalist phages, with baseplates formed from only five predicted subunits. This study evidences that multiple small mutations can be fixed into a sub-population of phage to provide a basis for phenotypic variation that we suggest could ultimately provide for a shift of virus properties, as an alternative evolutionary scenario to the major genetic events that result in the more well-studied evolutionary mechanism of phage mosaicism.