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Protein and nucleic acid synthesis in subcellular fractions of bacterial cells


, : Protein and nucleic acid synthesis in subcellular fractions of bacterial cells. Recent Progr Microbiol 7: 82-103

Starting with protoplasts, methods were devised to fractionate Escherichia coli cells centrifugally into 3 defined components; (a) soluble protein; (b) ribonucleoprotein particles; (c) protoplast membranes. The behavior of these fractions in situ leads to the conclusion that active synthesis of protein and ribo-nucleic acid occurs on the membranes. The other fractions are relatively inert. The in vitro behavior of these fractions with regard to protein synthesis is in agreement with this conclusion. The membrane fraction has more than 100 times the activity of the other 2 when measured in terms of incorporation of labeled amino acids into linkages stable to hot acid. It is the only fraction to exhibit any evidence that it can carry out the fabrication of complete protein molecules. The supplements required to observe optimal incorporation of amino acids in the membrane fraction suggested the presence of active nucleic acid metabolism. By including spermine in the incubation mixture it was possible to obtain extensive (10-fold) and reproducible net synthesis of polyribonucleotide. Optimal conditions for such synthesis require ribosidetriphosphates, adenosine triphosphate, Mn and a pH between 6.0 and 6.5. The reaction is inhibited by ortho- and pyrophosphates as well as by RNAase and DNAase. When supplemented with all 4 riboside triphosphates, the product formed has the properties of a polyribonucleotide strand containing a mixture of purine and pyrimidine bases.

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