Year of Award

2022

Document Type

Dissertation

Degree Type

Doctor of Philosophy (PhD)

Degree Name

Cellular, Molecular and Microbial Biology

Department or School/College

Division of Biological Sciences

Committee Chair

Patrick R. Secor

Commitee Members

Michael F. Minnick, Beverly J. Piggott, Brandon S. Cooper, Scott Wetzel, Stephen Lodmell

Keywords

bacteriophage, defense, infection, polyamine

Abstract

Cells are constantly monitoring their extracellular environment for danger signals that warn of imminent threats. When cells are killed by physical or other non-infectious injuries, they release concentrated intracellular molecules into the surrounding area. These danger-associated molecules alert other cells that danger is nearby. When cells are killed by pathogens, they release both damage-associated and pathogen-associated signals. Both eukaryotic and bacterial cells sense these molecules and make threat assessments of cellular injury. Here, I show that lysis of Pseudomonas aeruginosa and other bacterial species releases high concentrations of polyamines that are in turn taken up by surviving cells in a process mediated by Gac/Rsm and cyclic-di-GMP signaling. Intracellular polyamine levels spike in neighboring cells, with the duration of the high polyamine levels dictated by the infection status of the cell. In bacteriophage-infected cells, intracellular polyamine levels remain high, resulting in inhibition of bacteriophage genome replication. Linear DNA, whether in the form of packaged bacteriophage DNA or damage-induced linearized host DNA, was sufficient to induce intracellular polyamine accumulation, indicating that linear DNA is interpreted as a secondary danger signal. In addition, we discovered a group of N4-like bacteriophage that have evolved a means to disrupt intracellular polyamine accumulation and to allow for unencumbered phage replication. Because polyamines are ubiquitous to cellular life and Gac/Rsm and cyclic-di-GMP signaling are widespread amongst γ-proteobacteria, we propose that intracellular polyamine accumulation is a general strategy employed by bacteria to defend against bacteriophage infection and allow P. aeruginosa to make threat assessments of cellular injury.

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