Structural and transcriptional analyses of a purine nucleotide-binding protein from Pyrococcus furiosus: A component of a novel, membrane-bound multiprotein complex unique to this hyperthermophilic archaeon
The open-reading frame PF0895 in the genome of the hyperthermophilic archaeon, Pyrococcus furiosus, encodes a 206-residue protein (MR 23,152). The structure of the recombinant protein was solved by single isomorphous replacement with anomalous scattering (SIRAS) using a mercury derivative. It has been refined to 1.70 Å with a crystallographic R and Rfree values of 19.7% and 22.3%, respectively. The PF0895 structure is similar to those of the ATP binding cassettes observed in the ABC transporter family. However, bioinformatics and molecular analyses indicate that PF0895 is not part of the expected five-gene operon that encodes a typical prokaryotic solute-binding ABC transporter. Rather, transcriptional profiling data show that PF0895 is part of a novel four-gene operon (PF0895-PF0896-PF0897-PF0897.1) where only PF0895 has homologs in other organisms. Interestingly, from genome analysis, P. furiosus itself contains a second version of this complex, encoded by PF1090-PF1093. From the structural studies we can only conclude that one of the subunits of this novel membrane complex, PF0895, and its homolog PF1090, likely bind a purine nucleotide. PF0895 is therefore predicted to be part of a membrane-bound multiprotein complex unrelated to ABC transporters that is so far unique to P. furiosus. It appears to play a role in the stress response, as its expression is down regulated when the organism is subjected to cold-shock, where cells are transferred from 95°C, near the optimal growth temperature, to 72°C, near the minimal growth temperature. The related PF1090-containing operon is unaffected by cold-shock and is independently regulated. © 2007 Springer Science+Business Media B.V.
Gerwe, B; Kelley, L-LC; Dillard, BD; Lai, T; Liu, Z-J; Tempel, W; Chen, L; Habel, J; Lee, D; Jr, FEJ; Sugar, FJ; Richardson, JS; Richardson, DC; Newton, MG; Wang, B-C; Adams, MWW; Rose, JP
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