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  Bacterial‐like nitric oxide synthase in the haloalkaliphilic archaeon Natronomonas pharaonis

Orsini, S. S., James, K. L., Reyes, D. J., Couto-Rodriguez, R. L., Kolog Gulko, M., Witte, A., et al. (2020). Bacterial‐like nitric oxide synthase in the haloalkaliphilic archaeon Natronomonas pharaonis. MicrobiologyOpen, e1124. doi:10.1002/mbo3.1124.

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mbo3.1124.pdf (Publisher version), 4MB
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© 2020 The Authors.

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https://www.ncbi.nlm.nih.gov/bioproject/PRJNA630572 (Supplementary material)
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All DNA raw sequencing data files have been deposited to the National Center for Biotechnology Information's Sequence Read Archive (SRA), under BioProject PRJNA630572.

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 Creators:
Orsini, Silvia S.1, Author
James, Kimberly L.1, Author
Reyes, Destiny J.1, Author
Couto-Rodriguez, Ricardo L.1, Author
Kolog Gulko, Miriam2, Author              
Witte, Angela1, Author
Carroll, Ronan K.1, Author
Rice, Kelly C.1, Author
Affiliations:
1external, ou_persistent22              
2Oesterhelt, Dieter / Membrane Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565164              

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Free keywords: ARCHAEON HALOFERAX-VOLCANII; DEINOCOCCUS-RADIODURANS; PROTEIN; GENE; REVEALS; TRANSFORMATION; HALOARCHAEA; METABOLISM; SYSTEMS; OXYGENMicrobiology;
 Abstract: Bacterial nitric oxide (NO) synthases (bNOS) play diverse and important roles in microbial physiology, stress resistance, and virulence. Although bacterial and mammalian NOS enzymes have been well-characterized, comparatively little is known about the prevalence and function of NOS enzymes in Archaea. Analysis of archaeal genomes revealed that highly conserved bNOS homologs were restricted to members of the Halobacteria. Of these,Natronomonas pharaonisNOS (npNOS) was chosen for further characterization. NO production was confirmed in heterologously expressed His-tagged npNOS by coupling nitrite production from N-hydroxy-L-arginine in an H2O2-supported reaction. Additionally, thenosgene was successfully targeted and disrupted to create aNmn. pharaonis nosmutant by adapting an establishedNatrialba magadiitransformation protocol. Genome re-sequencing of this mutant revealed an additional frameshift in a putative cation-acetate symporter gene, which could contribute to altered acetate metabolism in thenosmutant. Inactivation ofNmn. pharaonis noswas also associated with several phenotypes congruent with bacterialnosmutants (altered growth, increased oxygen consumption, increased pigment, increased UV susceptibility), suggesting that NOS function may be conserved between bacteria and archaea. These studies are the first to describe genetic inactivation and characterization of aNmn. pharaonisgene and provides enhanced tools for probing its physiology.

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Language(s): eng - English
 Dates: 2020-11
 Publication Status: Published online
 Pages: 18
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000578237600001
DOI: 10.1002/mbo3.1124
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Title: MicrobiologyOpen
Source Genre: Journal
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Publ. Info: Malden, Mass. : Wiley
Pages: - Volume / Issue: - Sequence Number: e1124 Start / End Page: - Identifier: ISSN: 2045-8827
CoNE: https://pure.mpg.de/cone/journals/resource/2045-8827