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Creatine utilization as a sole nitrogen source in Pseudomonas putida KT2440 is transcriptionally regulated by CahR

Lauren A. Hinkel, Graham G. Willsey, Sean M. Lenahan, Korin Eckstrom, Kristin C. Schutz, View ORCID ProfileMatthew J. Wargo
doi: https://doi.org/10.1101/2021.11.02.466972
Lauren A. Hinkel
1Department of Microbiology and Molecular Genetics, University of Vermont Larner College of Medicine, Burlington, VT 05405
2Cellular, Molecular, and Biomedical Sciences Graduate Program, University of Vermont, Burlington, VT 05405
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Graham G. Willsey
1Department of Microbiology and Molecular Genetics, University of Vermont Larner College of Medicine, Burlington, VT 05405
2Cellular, Molecular, and Biomedical Sciences Graduate Program, University of Vermont, Burlington, VT 05405
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Sean M. Lenahan
2Cellular, Molecular, and Biomedical Sciences Graduate Program, University of Vermont, Burlington, VT 05405
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Korin Eckstrom
1Department of Microbiology and Molecular Genetics, University of Vermont Larner College of Medicine, Burlington, VT 05405
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Kristin C. Schutz
1Department of Microbiology and Molecular Genetics, University of Vermont Larner College of Medicine, Burlington, VT 05405
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Matthew J. Wargo
1Department of Microbiology and Molecular Genetics, University of Vermont Larner College of Medicine, Burlington, VT 05405
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  • ORCID record for Matthew J. Wargo
  • For correspondence: mwargo@uvm.edu
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ABSTRACT

Glutamine amidotransferase-1 domain-containing AraC-family transcriptional regulators (GATRs) are present in the genomes of many bacteria, including all Pseudomonas species. The involvement of several characterized GATRs in amine-containing compound metabolism has been determined, but the full scope of GATR ligands and regulatory networks are still unknown. Here, we characterize Pseudomonas putida’s detection of the animal-derived amine compound, creatine, a compound particularly enriched in muscle and ciliated cells by a creatine-specific GATR, PP_3665, here named CahR (Creatine amidohydrolase Regulator). cahR is necessary for transcription of the gene encoding creatinase (PP_3667/creA) in the presence of creatine and is critical for P. putida’s ability to utilize creatine as a sole source of nitrogen. The CahR/creatine regulon is small and electrophoretic mobility shift demonstrates strong and specific CahR binding only at the creA promoter, supporting the conclusion that much of the regulon is dependent on downstream metabolites. Phylogenetic analysis of creA orthologs associated with cahR orthologs highlights a strain distribution and organization supporting likely horizontal gene transfer, particularly evident within the genus Acinetobacter. This study identifies and characterizes the GATR that transcriptionally controls P. putida metabolism of creatine, broadening the scope of known GATR ligands and suggesting GATR diversification during evolution of metabolism for aliphatic nitrogen compounds.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE163362

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 02, 2021.
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Creatine utilization as a sole nitrogen source in Pseudomonas putida KT2440 is transcriptionally regulated by CahR
Lauren A. Hinkel, Graham G. Willsey, Sean M. Lenahan, Korin Eckstrom, Kristin C. Schutz, Matthew J. Wargo
bioRxiv 2021.11.02.466972; doi: https://doi.org/10.1101/2021.11.02.466972
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Creatine utilization as a sole nitrogen source in Pseudomonas putida KT2440 is transcriptionally regulated by CahR
Lauren A. Hinkel, Graham G. Willsey, Sean M. Lenahan, Korin Eckstrom, Kristin C. Schutz, Matthew J. Wargo
bioRxiv 2021.11.02.466972; doi: https://doi.org/10.1101/2021.11.02.466972

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