Dual role of a (p)ppGpp- and (p)ppApp-degrading enzyme in biofilm formation and interbacterial antagonism

Mol Microbiol. 2021 Jun;115(6):1339-1356. doi: 10.1111/mmi.14684. Epub 2021 Jan 25.


The guanosine nucleotide-based second messengers ppGpp and pppGpp (collectively: (p)ppGpp) enable adaptation of microorganisms to environmental changes and stress conditions. In contrast, the closely related adenosine nucleotides (p)ppApp are involved in type VI secretion system (T6SS)-mediated killing during bacterial competition. Long RelA-SpoT Homolog (RSH) enzymes regulate synthesis and degradation of (p)ppGpp (and potentially also (p)ppApp) through their synthetase and hydrolase domains, respectively. Small alarmone hydrolases (SAH) that consist of only a hydrolase domain are found in a variety of bacterial species, including the opportunistic human pathogen Pseudomonas aeruginosa. Here, we present the structure and mechanism of P. aeruginosa SAH showing that the enzyme promiscuously hydrolyses (p)ppGpp and (p)ppApp in a strictly manganese-dependent manner. While being dispensable for P. aeruginosa growth or swimming, swarming, and twitching motilities, its enzymatic activity is required for biofilm formation. Moreover, (p)ppApp-degradation by SAH provides protection against the T6SS (p)ppApp synthetase effector Tas1, suggesting that SAH enzymes can also serve as defense proteins during interbacterial competition.

Keywords: Pseudomonas aeruginosa; (p)ppApp; (p)ppGpp; biofilm formation; interbacterial competition; small alarmone hydrolase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine Nucleotides / metabolism*
  • Antibiosis / physiology*
  • Biofilms / growth & development
  • Gene Expression Regulation, Bacterial / genetics
  • Guanosine Pentaphosphate / metabolism*
  • N-Glycosyl Hydrolases / metabolism*
  • Pseudomonas aeruginosa / growth & development
  • Pseudomonas aeruginosa / metabolism*
  • Type VI Secretion Systems / metabolism*


  • Adenine Nucleotides
  • Type VI Secretion Systems
  • adenosine 3'-diphosphate 5'-diphosphate
  • Guanosine Pentaphosphate
  • N-Glycosyl Hydrolases