Glutamine versus Ammonia Utilization in the NAD Synthetase Family

NAD is a ubiquitous and essential metabolic redox cofactor which also functions as a substrate in certain regulatory pathways. The last step of NAD synthesis is the ATP-dependent amidation of deamido-NAD by NAD synthetase (NADS). Members of the NADS family are present in nearly all species across th...

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Main Authors:	De Ingeniis, Jessica, Kazanov, Marat D., Shatalin, Konstantin, Gelfand, Mikhail S., Osterman, Andrei L., Sorci, Leonardo
Format:	Online
Language:	English
Published:	Public Library of Science 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3376133/

id	pubmed-3376133
recordtype	oai_dc
spelling	pubmed-33761332012-06-20 Glutamine versus Ammonia Utilization in the NAD Synthetase Family De Ingeniis, Jessica Kazanov, Marat D. Shatalin, Konstantin Gelfand, Mikhail S. Osterman, Andrei L. Sorci, Leonardo Research Article NAD is a ubiquitous and essential metabolic redox cofactor which also functions as a substrate in certain regulatory pathways. The last step of NAD synthesis is the ATP-dependent amidation of deamido-NAD by NAD synthetase (NADS). Members of the NADS family are present in nearly all species across the three kingdoms of Life. In eukaryotic NADS, the core synthetase domain is fused with a nitrilase-like glutaminase domain supplying ammonia for the reaction. This two-domain NADS arrangement enabling the utilization of glutamine as nitrogen donor is also present in various bacterial lineages. However, many other bacterial members of NADS family do not contain a glutaminase domain, and they can utilize only ammonia (but not glutamine) in vitro. A single-domain NADS is also characteristic for nearly all Archaea, and its dependence on ammonia was demonstrated here for the representative enzyme from Methanocaldococcus jannaschi. However, a question about the actual in vivo nitrogen donor for single-domain members of the NADS family remained open: Is it glutamine hydrolyzed by a committed (but yet unknown) glutaminase subunit, as in most ATP-dependent amidotransferases, or free ammonia as in glutamine synthetase? Here we addressed this dilemma by combining evolutionary analysis of the NADS family with experimental characterization of two representative bacterial systems: a two-subunit NADS from Thermus thermophilus and a single-domain NADS from Salmonella typhimurium providing evidence that ammonia (and not glutamine) is the physiological substrate of a typical single-domain NADS. The latter represents the most likely ancestral form of NADS. The ability to utilize glutamine appears to have evolved via recruitment of a glutaminase subunit followed by domain fusion in an early branch of Bacteria. Further evolution of the NADS family included lineage-specific loss of one of the two alternative forms and horizontal gene transfer events. Lastly, we identified NADS structural elements associated with glutamine-utilizing capabilities. Public Library of Science 2012-06-15 /pmc/articles/PMC3376133/ /pubmed/22720044 http://dx.doi.org/10.1371/journal.pone.0039115 Text en De Ingeniis et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	De Ingeniis, Jessica Kazanov, Marat D. Shatalin, Konstantin Gelfand, Mikhail S. Osterman, Andrei L. Sorci, Leonardo
spellingShingle	De Ingeniis, Jessica Kazanov, Marat D. Shatalin, Konstantin Gelfand, Mikhail S. Osterman, Andrei L. Sorci, Leonardo Glutamine versus Ammonia Utilization in the NAD Synthetase Family
author_facet	De Ingeniis, Jessica Kazanov, Marat D. Shatalin, Konstantin Gelfand, Mikhail S. Osterman, Andrei L. Sorci, Leonardo
author_sort	De Ingeniis, Jessica
title	Glutamine versus Ammonia Utilization in the NAD Synthetase Family
title_short	Glutamine versus Ammonia Utilization in the NAD Synthetase Family
title_full	Glutamine versus Ammonia Utilization in the NAD Synthetase Family
title_fullStr	Glutamine versus Ammonia Utilization in the NAD Synthetase Family
title_full_unstemmed	Glutamine versus Ammonia Utilization in the NAD Synthetase Family
title_sort	glutamine versus ammonia utilization in the nad synthetase family
description	NAD is a ubiquitous and essential metabolic redox cofactor which also functions as a substrate in certain regulatory pathways. The last step of NAD synthesis is the ATP-dependent amidation of deamido-NAD by NAD synthetase (NADS). Members of the NADS family are present in nearly all species across the three kingdoms of Life. In eukaryotic NADS, the core synthetase domain is fused with a nitrilase-like glutaminase domain supplying ammonia for the reaction. This two-domain NADS arrangement enabling the utilization of glutamine as nitrogen donor is also present in various bacterial lineages. However, many other bacterial members of NADS family do not contain a glutaminase domain, and they can utilize only ammonia (but not glutamine) in vitro. A single-domain NADS is also characteristic for nearly all Archaea, and its dependence on ammonia was demonstrated here for the representative enzyme from Methanocaldococcus jannaschi. However, a question about the actual in vivo nitrogen donor for single-domain members of the NADS family remained open: Is it glutamine hydrolyzed by a committed (but yet unknown) glutaminase subunit, as in most ATP-dependent amidotransferases, or free ammonia as in glutamine synthetase? Here we addressed this dilemma by combining evolutionary analysis of the NADS family with experimental characterization of two representative bacterial systems: a two-subunit NADS from Thermus thermophilus and a single-domain NADS from Salmonella typhimurium providing evidence that ammonia (and not glutamine) is the physiological substrate of a typical single-domain NADS. The latter represents the most likely ancestral form of NADS. The ability to utilize glutamine appears to have evolved via recruitment of a glutaminase subunit followed by domain fusion in an early branch of Bacteria. Further evolution of the NADS family included lineage-specific loss of one of the two alternative forms and horizontal gene transfer events. Lastly, we identified NADS structural elements associated with glutamine-utilizing capabilities.
publisher	Public Library of Science
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3376133/
_version_	1611537120338378752

Glutamine versus Ammonia Utilization in the NAD Synthetase Family

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