Genomic and biochemical characterization of Acinetobacter podophage Petty reveals a novel lysis mechanism and tail-associated depolymerase activity

A. C. Hernandez-Morales, L. L. Lessor, T. L. Wood, D. Migl, E. M. Mijalis, J. Cahill, W. K. Russell, R. F. Young, J. J. Gill

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

The increased prevalence of drug-resistant, nosocomial Acinetobacter infections, particularly from pathogenic members of the Acinetobacter calcoaceticusbaumannii complex, necessitates the exploration of novel treatments such as phage therapy. In the present study, we characterized phage Petty, a novel podophage that infects multidrug-resistant Acinetobacter nosocomialis and Acinetobacter baumannii. Genome analysis reveals that phage Petty is a 40,431-bp ΦKMV-like phage, with a coding density of 92.2% and a G+C content of 42.3%. Interestingly, the lysis cassette encodes a class I holin and a single-subunit endolysin, but it lacks canonical spanins to disrupt the outer membrane. Analysis of other ΦKMV-like genomes revealed that spaninless lysis cassettes are a feature of phages infecting Acinetobacter within this subfamily of bacteriophages. The observed halo surrounding Petty's large clear plaques indicated the presence of a phage-encoded depolymerase capable of degrading capsular exopolysaccharides (EPS). The product of gene 39, a putative tail fiber, was hypothesized to possess depolymerase activity based on weak homology to previously reported phage tail fibers. The 101.4-kDa protein gene product 39 (gp39) was cloned and expressed, and its activity against Acinetobacter EPS in solution was determined. The enzyme degraded purified EPS from its host strain A. nosocomialis AU0783, reducing its viscosity, and generated reducing ends in solution, indicative of hydrolase activity. Given that the accessibility to cells within a biofilm is enhanced by degradation of EPS, phages with depolymerases may have enhanced diagnostic and therapeutic potential against drug-resistant Acinetobacter strains.

Original languageEnglish (US)
Article numbere01064-17
JournalJournal of virology
Volume92
Issue number6
DOIs
StatePublished - Mar 1 2018
Externally publishedYes

Keywords

  • Acinetobacter baumannii
  • Acinetobacter nosocomialis
  • Depolymerase
  • Genome analysis
  • Lysis
  • Phage therapy
  • Spanins

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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