Abstract
Protein-protein interactions among 960 Pyrococcus soluble proteins have been analysed by mammalian two-hybrid analysis and thirteen interactions between annotated and unannotated proteins detected.
Highlights
2,061 proteins of Pyrococcus horikoshii OT3, a hyperthermophilic archaeon, have been predicted from the recently completed genome sequence, the majority of proteins show no similarity to those from other organisms and are hypothetical proteins of unknown function
Using the SOSUI program [17], we examined the open reading frames (ORFs) to deduce which proteins included membrane-spanning sequences or signal peptide sequences; 410 clones were removed because they were predicted to code for membrane or secreted proteins
We found that the interacting pair of PH0487 and PH0490 have high similarity to the Bacillus subtilis chemotaxis proteins CheC and CheD, the genes for which are located adjacently on the Bacillus subtilis genome and compose an operon [23], suggesting that the ORFs for PH0487 and PH0490 are expressed in P. horikoshii OT3 as an operon and that their functions are similar to those of CheC and CheD
Summary
2,061 proteins of Pyrococcus horikoshii OT3, a hyperthermophilic archaeon, have been predicted from the recently completed genome sequence, the majority of proteins show no similarity to those from other organisms and are hypothetical proteins of unknown function. Because most proteins operate as parts of complexes to regulate biological processes, we systematically analyzed protein-protein interactions in Pyrococcus using the mammalian two-hybrid system to determine the function of the hypothetical proteins. Because most proteins operate as parts of complexes to regulate biological processes in cells or entire organisms, PPIs enable us to predict the functions of uncharacterized proteins through their associations with proteins of known function [3,4]. We used our mammalian two-hybrid system [11] to conduct a large-scale PPI analysis of the intracellular and soluble proteins of P. horikoshii OT3
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