Downregulation of Autolysin-Encoding Genes by Phage-Derived Lytic Proteins Inhibits Biofilm Formation in Staphylococcus aureus

Antimicrob Agents Chemother. 2017 Apr 24;61(5):e02724-16. doi: 10.1128/AAC.02724-16. Print 2017 May.

Abstract

Phage-derived lytic proteins are a promising alternative to conventional antimicrobials. One of their most interesting properties is that they do not readily select for resistant strains, which is likely due to the fact that their targets are essential for the viability of the bacterial cell. Moreover, genetic engineering allows the design of new "tailor-made" proteins that may exhibit improved antibacterial properties. One example of this is the chimeric protein CHAPSH3b, which consists of a catalytic domain from the virion-associated peptidoglycan hydrolase of phage vB_SauS-phiIPLA88 (HydH5) and the cell wall binding domain of lysostaphin. CHAPSH3b had previously shown the ability to kill Staphylococcus aureus cells. Here, we demonstrate that this lytic protein also has potential for the control of biofilm-embedded S. aureus cells. Additionally, subinhibitory doses of CHAPSH3b can decrease biofilm formation by some S. aureus strains. Transcriptional analysis revealed that exposure of S. aureus cells to this enzyme leads to the downregulation of several genes coding for bacterial autolysins. One of these proteins, namely, the major autolysin AtlA, is known to participate in staphylococcal biofilm development. Interestingly, an atl mutant strain did not display inhibition of biofilm development when grown at subinhibitory concentrations of CHAPSH3b, contrary to the observations made for the parental and complemented strains. Also, deletion of atl led to low-level resistance to CHAPSH3b and the endolysin LysH5. Overall, our results reveal new aspects that should be considered when designing new phage-derived lytic proteins aimed for antimicrobial applications.

Keywords: biofilms; novel antimicrobials; phage lytic proteins; transcriptional responses to antimicrobials.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Biofilms / drug effects
  • Biofilms / growth & development
  • Cell Wall / metabolism
  • Endopeptidases / metabolism
  • Lysostaphin / metabolism
  • N-Acetylmuramoyl-L-alanine Amidase / genetics*
  • Staphylococcus Phages / genetics*
  • Staphylococcus aureus / drug effects
  • Staphylococcus aureus / genetics
  • Staphylococcus aureus / metabolism*
  • Staphylococcus aureus / virology*
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / pharmacology*

Substances

  • Anti-Bacterial Agents
  • Viral Fusion Proteins
  • Endopeptidases
  • Lysostaphin
  • endolysin
  • N-Acetylmuramoyl-L-alanine Amidase