CHANGES IN SUBCELLULAR COMPONENTS OF YEAST AS RELATED TO NUCLEOTIDE RELEASE FROM THE CELLS INCUBATED IN CITRATE BUFFER

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It was found that when intact cells of a yeast, Saccharomyces celevisiae 'Yebisu', were incubated in 0.08M citrate buffer (pH 6.0) containing 2 per cent glucose, nucleotides were released in the medium. In this connection, experiments have been carried out to elucidate biochemical changes in subcellular structure of such cells. Microscopic observation showed that the longer the duration of incubation of the cells in the citrate buffer, the more marked becomes the granulous appearance of the cytoplasm. Among various subcellular fractions of freshly disrupted cells, the highest content in nucleic acid was found in the cell membrane fraction and in the small granule fraction. The nucleic acid content in the former fraction decreased markedly, even after a short period of incubation with citrate, accompanied by an abundant release of nucleotides. In contrast, the nucleic acid content in the small granule fraction scarcely changed. Continued incubation with citrate, however, caused a decrease of nucleic acid content also in this fraction. In this case, also the extracellular release of amino acids increased and a partial loss of viability of the cells was observed. Ultracentrifugal analysis showed that the sedimentation pattern of the small granule fraction, consisting of an 80S (major) and a 40S (minor) component, did not change on incubation with citrate.
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