Research on bacterial nnodel systems has established much of our understanding of basic molecular biology. Extrapolations from bacterial systems to eukaryotic molecular biology and human health issues are however inherently limited by the lack of bacterial homologues of many components conserved in eukaryotic cells. Fortunately, Archaea do have homologues of many of these "eukaryotic" components and research with such simpler archaeal systems can therefore be legitimately extrapolated to eukaryotic/human cellular and molecular biology. The archaeal machineries, for example, that catalyze DNA replication and repair, transcription, transcript and protein processing, translation initiation, cytokinesis and cell division are all far simpler than their eukaryotic counterparts, but most of their components are proteins that have well-conserved structural and likely functional eukaryotic homologues. For experimental research, the attraction of these much simpler but eukaryote-homologous prokaryotic systems is obvious and palpable, but with very limited genetics and no archaeal model system, access of the research community to the advantages and opportunities afforded by the Archaea has been effectively blocked. We have recently established the genetic techniques needed for Thermococcus kodakarensis (T.k.) and, as solicited by the PA-07-457 request "...to develop and distribute genetic and genomic resources for emerging non-mammalian model organisms" the project proposed will generate the genomic resources needed to fully establish T.k. as a facile archaeal model system. Specifically, every protein-encoding gene will be Individually cloned and also modified to add a hemagglutinin (HA) epitope for Identification and a Hls6-afflnlty-tag for purification In sequence-verified plasmid libraries. T.k. strain libraries will also be constructed with every non-essential open reading frame (ORF) Individually deleted, and every genomic ORF HA-His6 extended. With the T.k. genetic techniques established and these genomic resources constructed, the first archaeal model system will be available. This will facilitate, expedite and encourage Investigations ofthe many archaeal cellular and molecular biology svstems that are homologues of eukarvotic/human systems and so valid models for healthcare research.

Public Health Relevance

Archaea offer the research advantages of far less complexity but close structural and functional homology and so direct relevance to human cellular and molecular biology. This project will establish the first archaeal model system, supported by comprehensive strain and gene libraries. This will be a new and unique genetic resource that provides the biomedical research community with direct and Immediate access to the many archaeal systems that are legitimate experimental models for human biology and for biomedical research.

Agency
National Institute of Health (NIH)
Type
Resource-Related Research Projects (R24)
Project #
5R24GM098176-04
Application #
8720014
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Willis, Kristine Amalee
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Ohio State University
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
City
Columbus
State
OH
Country
United States
Zip Code
43210
Jäger, Dominik; Förstner, Konrad U; Sharma, Cynthia M et al. (2014) Primary transcriptome map of the hyperthermophilic archaeon Thermococcus kodakarensis. BMC Genomics 15:684
Li, Zhuo; Huang, Richard Y-C; Yopp, Daniel C et al. (2014) A novel mechanism for regulating the activity of proliferating cell nuclear antigen by a small protein. Nucleic Acids Res 42:5776-89
Cubonova, Lubomira; Richardson, Tomas; Burkhart, Brett W et al. (2013) Archaeal DNA polymerase D but not DNA polymerase B is required for genome replication in Thermococcus kodakarensis. J Bacteriol 195:2322-8
Nalabothula, Narasimharao; Xi, Liqun; Bhattacharyya, Sucharita et al. (2013) Archaeal nucleosome positioning in vivo and in vitro is directed by primary sequence motifs. BMC Genomics 14:391