Substrate identification of putative NCS1 and NCS2 nucleobase transporters in Pseudomonas aeruginosa

Corey Kennelly, Arthur Prindle*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Pseudomonas aeruginosa is an opportunistic pathogen that can salvage nucleobases from the environment to conserve nutrients that would otherwise be spent on de novo nucleotide biosynthesis. However, little is known regarding the substrate specificity of the 13 putative nucleobase transporters in P. aeruginosa. Here, using a combination of genetic and chemical approaches, we report substrate identifications for 10 putative nucleobase transporters in P. aeruginosa. Specifically, we individually expressed each transporter in a genetic background lacking all 13 putative nucleobase transporters and quantified growth on a panel of 10 nucleobases as sole nitrogen sources. We confirmed these expression-based substrate identifications using targeted genetic knockouts. In a complementary approach, we utilized four toxic nucleobase antimetabolites to characterize antimicrobial activity in these same strains. We identified the sole allantoin transporter as well as transporters for guanine, xanthine, uric acid, cytosine, thymine, uracil, and dihydrouracil. Furthermore, we associated at least five nucleobase transporters with hypoxanthine, which has been recently reported to be an antibiofilm cue in P. aeruginosa. These results provide an initial characterization of the putative nucleobase transporters in P. aeruginosa, significantly advancing our understanding of nucleobase transport in this clinically relevant organism.

Original languageEnglish (US)
JournalmBio
Volume15
Issue number12
DOIs
StatePublished - Dec 2024

Funding

We thank Blake Everett for comments during the writing of this article and members of the Prindle Lab for their support. We thank Colin Manoil for MPAO1 and Kelly Bachta and Alan Hauser for S17-1 \u03BBpir E. coli, pEXG2, and pPSV37. We are grateful for the generous support from the David and Lucile Packard Foundation (2018\u201368055), the Army Research Office (W911NF-19-1-0136), Pew Charitable Trusts (2019-A-06953), the National Science Foundation (NSF 2239567), the National Institutes of Health (1R35GM147170-01), and a Dr. John N. Nicholson Fellowship from Northwestern University.

Keywords

  • nucleobase
  • Pseudomonas aeruginosa
  • purines
  • pyrimidines
  • transporters

ASJC Scopus subject areas

  • Microbiology
  • Virology

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