Methanococcus maripaludis is an ideal model organism for the study of transcriptional regulation in the Archaea. Through the study of a nitrogen assimilation regulon, novel aspects of transcriptional regulation are coming to light. A complete system has been identified for detailed study: a novel repressor protein NrpR, an inducer (2-oxoglutarate) that modulates NrpR activity, three regulated promoter regions, and three different nitrogen sources that result in different regulatory states. Each promoter region contains a different configuration of operator sites and has a different regulatory outcome. A robust set of genetic tools allows for testing the effects of various manipulations on regulatory outcome in vivo. The system provides the opportunity to understand in detail a particular system of transcriptional regulation in the Archaea. Experiments will determine the subunit structure of the NrpR-DNA complexes, the binding properties of NrpR to each promoter region, how 2-oxoglutarate modulates binding, and how regulatory outcome is determined. In addition genetic studies will be carried out to identify the domain of NrpR that binds 2- oxoglutarate and the domain that mediates multimer formation. Knowledge of regulation in the Archaea will be greatly increased. Comparison with known mechanisms in the Bacteria and Eukarya may reveal fundamental similarities or differences in regulatory mechanisms. Archaeal transcription is a good model for eukaryotic transcription due to its distinct relatedness but relative simplicity. These studies in Archaea have wide importance, as regulatory mechanisms are central to human development and disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM055255-05A1
Application #
6721586
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Program Officer
Anderson, James J
Project Start
1998-05-01
Project End
2007-12-31
Budget Start
2004-01-01
Budget End
2004-12-31
Support Year
5
Fiscal Year
2004
Total Cost
$260,207
Indirect Cost
Name
University of Washington
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Lie, Thomas J; Hendrickson, Erik L; Niess, Ulf M et al. (2010) Overlapping repressor binding sites regulate expression of the Methanococcus maripaludis glnK(1) operon. Mol Microbiol 75:755-62
Wisedchaisri, Goragot; Dranow, David M; Lie, Thomas J et al. (2010) Structural underpinnings of nitrogen regulation by the prototypical nitrogen-responsive transcriptional factor NrpR. Structure 18:1512-21
Xia, Qiangwei; Wang, Tiansong; Hendrickson, Erik L et al. (2009) Quantitative proteomics of nutrient limitation in the hydrogenotrophic methanogen Methanococcus maripaludis. BMC Microbiol 9:149
Lie, Thomas J; Leigh, John A (2007) Genetic screen for regulatory mutations in Methanococcus maripaludis and its use in identification of induction-deficient mutants of the euryarchaeal repressor NrpR. Appl Environ Microbiol 73:6595-600
Leigh, John A; Dodsworth, Jeremy A (2007) Nitrogen regulation in bacteria and archaea. Annu Rev Microbiol 61:349-77
Lie, Thomas J; Dodsworth, Jeremy A; Nickle, David C et al. (2007) Diverse homologues of the archaeal repressor NrpR function similarly in nitrogen regulation. FEMS Microbiol Lett 271:281-8
Moore, Brian C; Leigh, John A (2005) Markerless mutagenesis in Methanococcus maripaludis demonstrates roles for alanine dehydrogenase, alanine racemase, and alanine permease. J Bacteriol 187:972-9
Lie, Thomas J; Wood, Gwendolyn E; Leigh, John A (2005) Regulation of nif expression in Methanococcus maripaludis: roles of the euryarchaeal repressor NrpR, 2-oxoglutarate, and two operators. J Biol Chem 280:5236-41
Wood, Gwendolyn E; Haydock, Andrew K; Leigh, John A (2003) Function and regulation of the formate dehydrogenase genes of the methanogenic archaeon Methanococcus maripaludis. J Bacteriol 185:2548-54
Lie, Thomas J; Leigh, John A (2003) A novel repressor of nif and glnA expression in the methanogenic archaeon Methanococcus maripaludis. Mol Microbiol 47:235-46

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