Phage-encoded cationic antimicrobial peptide used for outer membrane disruption in lysis Institutional Repository Document uri icon

abstract

  • AbstractSpanins are required for the last step in bacteriophage lysis: the disruption of the outer membrane. Bioinformatic analysis has shown that ~15% of phages lack a spanin gene, which suggests an alternate mechanism of outer membrane disruption. To address this, we selected virulent podophage KT as a spaninless exemplar and tested KT genes for outer membrane disruption during lysis. Hypothetical novel gene 28 causes outer membrane disruption when co-expressed with KT lysis genes and complements the lysis defect of a spanin mutant. Gp28 is a 56 aa cationic peptide with predicted amphipathic helical structure and is associated with the particulate fraction after lysis. Urea and KCl washes did not release gp28 from the particulate, suggesting a strong hydrophobic interaction with the membrane. Super high-resolution microscopy supports a primarily outer membrane localization for the peptide. Additionally, holin function is not required for gp28-mediated lysis. Gp28 is similar in size, charge, predicted fold, and membrane association to the human cathelicidin antimicrobial peptide LL-37. In standard assays to measure bactericidal and inhibitory effects of antimicrobial peptides on bacterial cells, synthesized gp28 performed equivalently to LL-37. The studies presented here suggest that KT Gp28 disrupts bacterial outer membranes during lysis in a manner akin to antimicrobial peptides.SignificanceHere we provide evidence that KT produces an antimicrobial peptide for outer membrane disruption during lysis. The disruptin is a new paradigm for phage lysis, and has no similarities to other known lysis genes. Many mechanisms have been proposed for the function of antimicrobial peptides, however there is not a consensus on the molecular basis of membrane disruption. Additionally, there is no established genetic selection system to support such studies. Therefore, the KT disruptin may represent the first genetically tractable antimicrobial peptide.

altmetric score

  • 26.45

author list (cited authors)

  • Holt, A., Cahill, J., Ramsey, J., OLeary, C., Moreland, R., Martin, C., ... Young, R.

citation count

  • 4

complete list of authors

  • Holt, Ashley||Cahill, Jesse||Ramsey, Jolene||O‚ÄôLeary, Chandler||Moreland, Russell||Martin, Cody||Galbadage, Don Thushara||Sharan, Riti||Sule, Preeti||Bettridge, Kelsey||Xiao, Jie||Cirillo, Jeffrey||Young, Ryland

Book Title

  • bioRxiv

publication date

  • January 2019