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A Putative ABC Transporter, HatABCDE, Is among Molecular Determinants of Pyomelanin Production in Pseudomonas aeruginosa

Hunter, Ryan C. and Newman, Dianne K. (2010) A Putative ABC Transporter, HatABCDE, Is among Molecular Determinants of Pyomelanin Production in Pseudomonas aeruginosa. Journal of Bacteriology, 192 (22). pp. 5962-5971. ISSN 0021-9193. PMCID PMC2976449.

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Pyomelanin overproduction is a common phenotype among Pseudomonas aeruginosa isolates recovered from cystic fibrosis and urinary tract infections. Its prevalence suggests that it contributes to the persistence of the producing microbial community, yet little is known about the mechanisms of its production. Using transposon mutagenesis, we identified factors that contribute to melanogenesis in a clinical isolate of P. aeruginosa. In addition to two enzymes already known to be involved in its biosynthesis (homogentisate dioxygenase and hydroxyphenylpyruvate dioxygenase), we identified 26 genes that encode regulatory, metabolic, transport, and hypothetical proteins that contribute to the production of homogentisic acid (HGA), the monomeric precursor of pyomelanin. One of these, PA14_57880, was independently identified four times and is predicted to encode the ATP-binding cassette of an ABC transporter homologous to proteins in Pseudomonas putida responsible for the extrusion of organic solvents from the cytosol. Quantification of HGA production by P. aeruginosa PA14 strains missing the predicted subcomponents of this transporter confirmed its role in HGA production: mutants unable to produce the ATP-binding cassette (PA14_57880) or the permease (PA14_57870) produced substantially less extracellular HGA after growth for 20 h than the parental strain. In these mutants, concurrent accumulation of intracellular HGA was observed. In addition, quantitative real-time PCR revealed that intracellular accumulation of HGA elicits upregulation of these transport genes. Based on their involvement in homogentisic acid transport, we rename the genes of this operon hatABCDE.

Item Type:Article
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URLURL TypeDescription CentralArticle
Hunter, Ryan C.0000-0003-3841-1676
Newman, Dianne K.0000-0003-1647-1918
Additional Information:© 2010 American Society for Microbiology. Received 27 August 2010; Accepted 10 September 2010. Published ahead of print on 24 September 2010. R.C.H. was supported by a Canadian Cystic Fibrosis Foundation Postdoctoral Fellowship, and D.K.N. is an investigator for the Howard Hughes Medical Institute. We thank Max Schöbert (Hannover Medical School) for providing the pyomelanin-overpoducing isolate and Nicole Denisco, Maureen Coleman, Paula Welander, Lars Dietrich, and Alexa Price-Whelan for technical assistance and helpful discussions.
Funding AgencyGrant Number
Canadian Cystic Fibrosis FoundationUNSPECIFIED
Issue or Number:22
PubMed Central ID:PMC2976449
Record Number:CaltechAUTHORS:20101103-111826151
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:20654
Deposited By: Jason Perez
Deposited On:03 Nov 2010 20:12
Last Modified:09 Mar 2020 13:18

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