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PLoS One. 2015 Dec 15;10(12):e0144514. doi: 10.1371/journal.pone.0144514. eCollection 2015.

Genomic and Gene-Expression Comparisons among Phage-Resistant Type-IV Pilus Mutants of Pseudomonas syringae pathovar phaseolicola.

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School of Natural Sciences, University of California Merced, Merced, 95343, CA, United States of America.
Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT 06520, United States of America.
Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario, M5S 3B2, Canada.
Centre for the Analysis of Genome Evolution & Function, University of Toronto, Ontario, M5S 3B2, Canada.
Department of Biostatistics, Yale School of Public Health, New Haven, CT 06520, United States of America.
Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT 06520, United States of America.
Program in Microbiology, Yale University, New Haven, CT 06520, United States of America.


Pseudomonas syringae pv. phaseolicola (Pph) is a significant bacterial pathogen of agricultural crops, and phage Φ6 and other members of the dsRNA virus family Cystoviridae undergo lytic (virulent) infection of Pph, using the type IV pilus as the initial site of cellular attachment. Despite the popularity of Pph/phage Φ6 as a model system in evolutionary biology, Pph resistance to phage Φ6 remains poorly characterized. To investigate differences between phage Φ6 resistant Pph strains, we examined genomic and gene expression variation among three bacterial genotypes that differ in the number of type IV pili expressed per cell: ordinary (wild-type), non-piliated, and super-piliated. Genome sequencing of non-piliated and super-piliated Pph identified few mutations that separate these genotypes from wild type Pph--and none present in genes known to be directly involved in type IV pilus expression. Expression analysis revealed that 81.1% of gene ontology (GO) terms up-regulated in the non-piliated strain were down-regulated in the super-piliated strain. This differential expression is particularly prevalent in genes associated with respiration--specifically genes in the tricarboxylic acid cycle (TCA) cycle, aerobic respiration, and acetyl-CoA metabolism. The expression patterns of the TCA pathway appear to be generally up and down-regulated, in non-piliated and super-piliated Pph respectively. As pilus retraction is mediated by an ATP motor, loss of retraction ability might lead to a lower energy draw on the bacterial cell, leading to a different energy balance than wild type. The lower metabolic rate of the super-piliated strain is potentially a result of its loss of ability to retract.

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