156. Whole Genome Sequence and Structural Proteomics of Pyrobaculum aerophilum 

Sorel Fitz-Gibbon¹, Ung-Jin Kim², Heidi Ladner¹, Elizabeth Conzevoy¹, Gigi Park², Karl Stetter³, Jeffrey H. Miller¹, and Melvin I. Simon^2 
1Department of Microbiology and Molecular Biology Institute, University of California, Los Angeles, California; ²Biology, California Institute of Technology, Pasadena, California; and ³University of Regensberg, Germany 
sorel@mbi.ucla.edu 

Pyrobaculum aerophilum is a hyperthermophilic archaeon, isolated from a boiling marine water hole, that is capable of growth at 104C. This microorganism can grow microaerobically, unlike most of its thermophilic relatives, making it amenable to a variety of experimental manipulations and a candidate as a model organism for studying archaeal and thermophilic microbiology. We have sequenced the entire genome using a random shotgun approach (3.5X genomic coverage) followed by oligonucleotide primer directed sequencing, guided by our fosmid map. The 2.2 Mb genome codes for more than 2000 proteins, 30% of which have been identified by their sequence similarities to proteins of known function. Only 15% of the Pyrobaculum aerophilum proteins have related high resolution structures. In collaboration with the DOE/UCLA Laboratories and Los Alamos National Laboratories we have initiated a project to express and purify proteins for structure determination by NMR or xray diffraction. The three dimensional structures of the Pyrobaculum aerophilum proteins will give one the power to understand and manipulate protein function and are crucial to fully exploiting the information in the genome. At this time several proteins have been cloned, expressed in E.coli, purified and crystals which diffract to high resolution have been obtained.