The degree of barotolerance exhibited by Pseudomonas fluorescens and Pseudomonas bathycetes in vitro polyphenylalanine-synthesizing systems can be modified by altering the concentrations of specific ions in the reaction mixture. Hybrid-protein-synthesizing systems, utilizing all the possible S-100 supernatant fluid and ribosome combinations from Escherichia coli, P. fluorescens, and P. bathycetes, were tested for barotolerance under conditions of low (16 mM Mg2+ plus 0 mM Na+) and high (150 mM Na+ plus 60 mM Mg2+) ion concentrations. The results reveal that barotolerant synthesis is a characteristic determined by the origin of the ribosome. Systems utilizing E. coli ribosomes are barosensitive at both low and high ion concentrations, P. fluorescens ribosomes barotolerant under both conditions, and P. bathycetes ribosomes barosensitive at low and barotolerant at high ion concentrations. Therefore certain concentrations of specific ions will increase barotolerance, but only if the ribosomes are capable of functioning at high pressures.Protein synthesis in Escherichia coli has been shown to be extremely sensitive to application of high hydrostatic pressures (1-3, 6-8, 14, 15). The primary inhibitory effect of high pressure on the translation process has been postulated to involve the ribosome directly, by possibly inhibiting the binding of amino acyl transfer ribonucleic acid (RNA) to the ribosome-messenger RNA complex or by interfering with translocation. It has been recently reported that protein synthesis in Pseudomonas fluorescens, whether in whole-cell or cellextract preparations, displays a marked barotolerance, i.e., a resistance to the inhibitory effects of high pressure (16). The availability of both a barosensitive E. coli system and a barotolerant P. fluorescens system thus allowed for a series of experiments involving interchanges of ribosomes and supernatant factors in polyuridylate [poly(U)]-directed, polyphenylalaninesynthesizing preparations. Such experiments showed clearly that barotolerance in P. fluorescens was associated with the ribosomes rather than the soluble components of the system (13). Additional manipulation of the 50S and 30S ribosomal subunits in interchange experiments revealed that resistance to inhibition by pressure was associated with the 30S ribosomal subunit of P. fluorescens (18).Our initial concept was to compare the effects of pressure on protein synthesis in E. coli and in Pseudomonas bathycetes, a deep-sea isolate from the Marianas Trench where the pressure exceeds 1,000 atm. P. fluorescens was selected as a terrestrial pseudomonad for limited generic comparison with P. bathycetes. P. bathycetes grows at higher pressures (16) and protein synthesis was found to be relatively resistant (when compared with E. coli) to high pressure in whole-cell preparations (16). Surprisingly, this barotolerant characteristic was lost in cellextract preparations. That is, poly(U)-directed synthesizing systems of P. bathycetes showed the same sensitivity to pressure as that shown by E. coli p...