Uptake of alpha-aminoisobutyric acid (AIB) by a leucine-tyrosine auxotroph of a thermophilic microorganism starved for leucine was studied. AIB was taken up by the cells against a substantial concentration gradient (300:1) and was present there in a free and unchanged form. Various energy inhibitors and sulfhydryl reagents strongly inhibited the accumulation of AIB. AIB uptake obeyed saturation kinetics, and the Lineweaver-Burk plot is characterized by a biphasic curve. AIB most probably shares a common transport system(s) with alanine, serine, and glycine. A mutant defective in l-alanine uptake was isolated by using the suicide effect due to accumulation of the tritiated substrate. The mutant also exhibited impaired transport activity towards AIB, glycine, and l-serine, but not to phenylalanine or valine. The transport of AIB, glycine, l-alanine, and l-serine was induced by d-alanine (5 x 10(-3) M) during growth in a succinate- and ammonia-containing medium. De novo protein synthesis was required for the induction of AIB transport; the induction was inhibited when growth occurred in glucose-containing media. The apparent differential rate of synthesis of the AIB transport system was decreased considerably in glucose-grown cells as compared to succinate-grown cells. A common genetic basis of either the regulatory or structural nature for the transport of AIB, alanine, glycine, and serine in a thermophilic microorganism is suggested.