Polynucleotide Phosphorylysis and Synthesis by Lysed Cells of Micrococcus Iysodeikticus

Abstract

AQUEOUS extracts of Micrococcus lysodeikticus possess a highly active enzyme capable of polymerizing mononucleotides reversibly¹. In addition, the bacterial ribonucleic acid is readily phosphorylysed by this enzyme in the presence of inorganic phosphate. Because of these facts, we have attempted in a few preliminary experiments to evaluate the part this enzyme system may play in modifying the properties or content of bacterial cell ribonucleic acid during lysis of the cells and extraction procedures. In these experiments we have measured the acid-insoluble and acid-soluble fractions extractable from the cells during and after lysis in tris buffer, phosphate buffer, and in the presence of adenosine diphosphate in tris buffer, all experiments at pH 9.5. The cells were harvested from submerged 48-hr. cultures² and washed three times in 0.5 per cent sodium chloride. The concentration of cells was selected empirically by choosing a value which would, after complete lysis, yield an optical density of approximately 1.0 at 260 mµ for the acid-insoluble material when dissolved in 3 ml. of tris solution, pH 8.0, 0.1 M.