John N. Reeve

reeve.2@osu.edu

Rod Sharp Professor of Microbiology
Department Chair
Ph.D., University of British Columbia, 1971

Molecular biology of archaea \ molecular adaptations to extreme environments.

Archaea are prokaryotes, many of which have unusual life-styles and unique metabolic abilities. Some live in very extreme environments, such as the boiling water surrounding volcanic vents, in saturated salt, highly acidic and alkaline lakes, and in the complete absence of oxygen. Our research interests are in the molecular biology of gene expression in two thermophilic Archaea, a methanogen (Methanothermobacter thermautotrophicus) that grows optimally at 65°C, and a fermentative heterotroph (Thermococcus kodakarensis) that grows optimally at 85°C. Research has established that the DNA, RNA and protein synthesizing machineries in Archaea are simpler but apparently closely related and likely ancestral to their counterparts in the eukaryotic nucleus. Our experiments combine in vivo genetics and in vitro biochemistry to identify the components and determine the mechanics and regulation of these archaeal molecular biology machines. Some Archaea also have histones, small DNA-binding proteins that are homologues of the nucleosome core histones that compact nuclear DNA into chromatin and provide the targets for epigenetic regulation eukaryotic chromosomal DNA. We are determining the structures, functions and evolutionary relationships of archaeal histones to their eukaryotic counterparts.

Methane (natural gas) production from biomass is an attractive alternative to ethanol as a renewable source of energy. Biogas (~60% methane) is generated from waste biomass and used locally as a fuel in many regions of the world, and methane is also generated in all urban and industrial waste-treatment facilities, and in land-fills, but little of this methane is currently recovered as fuel. Our research on gene expression in a methanogen also has economic value therefore as the knowledge gained should help facilitate genetic engineering to improve large-scale methane production for energy, and waste management. Research on hyperthermophilic Archaea also has an applied biotechnology component as many of the heat-resistant enzymes present in these species have attractive features as industrial catalysts. Our research should help develop heat-resistant archaeal enzymes for commercial uses.

Xie, Y. and Reeve, J.N. 2004. Transcription by an archaeal RNA polymerase is slowed but not blocked by an archaeal nucleosome. J. Bacteriol. 186:3492-3498.

Xie, Y. and Reeve, J.N. 2004. Transcription by Methanothermobacter thermautotrophicus RNA polymerase in vitro releases archaeal transcription factor B but not TATA-box binding protein from the template DNA. J. Bacteriol. 186:6306-6310.

Reeve, J.N., Bailey, K.A., Li, W.T., Marc, F., Sandman, K. and Soares, D.J. 2004. Archaeal histones: structures, stability and DNA binding. Biochem. Soc. Trans. 32:227-230.

Cubonová, L., Sandman, K., Hallam, S.J., DeLong, E.F., and Reeve, J.N. 2005. Histones in Crenarchaea. J. Bacteriol. 187:5482-5485.

Reeve, J.N., and Schmitz, R.A. 2005. Biology, biochemistry and the molecular machinery of Archaea. Curr. Opin Microbiol. 8:627-629.

Sandman, K., and Reeve, J.N. 2005. Archaeal chromatin proteins. Curr. Opin Microbiol. 8:656-661.

Shin, J-H., Reeve, J.N., and Kelman, Z. 2005. Use of a restriction enzyme-digested PCR product as substrate for helicase assays. Nucl. Acids Res. 33:e8.

Xie, Y., and Reeve, J.N. 2005. Regulation of tryptophan operon expression in the Archaeon Methanothermobacter thermautotrophicus. J. Bacteriol. 187:6419-6429.

Sandman, K., and Reeve, J.N. 2006. Archaeal histones and the origin of the histone fold. Curr. Opin. Microbiol. 9:520-525.

Santangelo, T.J. and Reeve, J.N. 2006. Archaeal RNA polymerase is sensitive to intrinsic termination directed by transcribed and remote sequences. J. Mol. Biol. 355:196-210.

Shin, J-H., Santangelo, T.J., Xie, Y., Reeve, J.N., and Kelman, Z. 2007. Archaeal MCM helicase can unwind DNA bound by archaeal histones and transcription factors. J. Biol Chem. 282:4908-4915.

Reeve, J.N., Xie, Y., and Santangelo, T.J. 2007. Regulation of transcription initiation and termination in Archaea. Proc Int. Symp Extremophiles and their Applications. Japan Agency for Marine-Earth Science and Technology. pp 2-8.

Reeve, J.N., and Sandman, K. 2007. Chromatin and regulation. In Archaea: Evolution, Physiology, and Molecular Biology. Eds Garrett, R., and Klenk, H-P. Blackwell Publ., Oxford UK. pp 147-158.

Santangelo, T.J., Cubonova, L., James, C.L., and Reeve, J.N. 2007. TFB1 or TFB2 is sufficient for Thermococcus kodakarensis viability and basal transcription in vitro. J. Mol. Biol. 367:344-357.

French, S.L, Santangelo, T.J, Beyer, A.L., and Reeve, J.N. 2007. Transcription and translation are coupled in Archaea. Mol. Biol. Evol. 24:893-895.

Cubonova, L, Sandman, K., Karr, E.A., Cochran, A.J., and Reeve, J.N. 2007. Spontaneous mutagenesis of trpY and mutational analysis of the TrpY archaeal transcription regulator. J. Bacteriol. 189:4338-4342.

Samson, R., and Reeve, J.N. 2007. DNA binding proteins and chromatin. In Archaea: Molecular and Cellular Biology. Ed. Cavicchioli, R. ASM Press, Washington, DC. USA. pp. 110-119.