The mutant T44(lambda) of Escherichia coli K12, grown in the presence of adenine, develops an increased tolerance to streptomycin. In cultures grown on streptomycin, the ts character (tif) may temporarily be suppressed but, on further transfer, both the temperature-sensitive phenotype and streptomycin tolerance disappear. In a cell-free system, the relative efficiency of translation of MS2 and poly U messenger RNAs was, respectively, 75 and 50% lower in extracts from cultures grown at 37 degrees with adenine than in extracts from 30 degrees cultures. Similar results were obtained when adenine was added in vitro to an extract from a culture grown at 37 degrees in the absence of adenine, using MS2 RNA as messenger. Moreover, the 37 degrees extracts showed a much lower misincorporation of isoleucine into polyphenylalanine in the poly U system. In addition, the Mg++ concentration required for optimal translational acitvity was higher for the 37 degrees than for the 30 degrees extracts. Extracts from a culture grown in L medium at 37 degrees or from a tif-/F'tif+ merodiploid grown at 37 degrees with adenine behaved similarly to that from the 30 degrees culture when poly U was used as messenger RNA. It is suggested that the tif+ gene product may play a regulatory role in ribosomal function and the pleiotropic nature of the tif-1 mutation could be due to impairment of translational activity augmented by elevated temperature or by adenine.