1. The stability of NNMG with respect to pH was investigated. A maximum, with a half-life of 40 hours, was found in phosphate-citric acid buffer at pH 5. The stability decreases quickly with increasing concentration of hydroxyl ions. At the optimal mutation rate (pH 6, 37°, and shaking) the half-live is only 14,4 hours.2. NNMG uptake (using labelled NNMG) and mutation rate in the range pH 3,5 — 8,0 show an optimum at pH 6,0/6,5 for both Escherichia coli B and the red ad- mutant E 188 of Saccharomyces cerevisiae. The pH-dependence of NNMG uptake and mutation rate is similar.3. The methylation of nucleic acids by means of [3H]-methyl-NNMG or [14C] -methyl-NNMG in phosphate-citric acid at 37° while shaking increases with the growing concentration of hydroxyl ions. At the optimal mutation rate, however, NNMG methylates relatively poor.4. Incorporation of radioactive NNMG into Escherichia coli B results in labelling of nucleic acids and proteins. The labelling of DNA shows an optimum at pH 6. The extent of methylation was found to be higher in vivo than in vitro.5. In the presence of cysteine or β-mercaptoethanol nucleic acids are methylated more intensively than in the cysteine-free control experiment. After one hour the methylation was about twentyfold higher in the presence of cysteine or β-mercaptoethanol compared to the cysteine-free control experiment. The methylation in presence of SH-compounds shows a maximum at pH 6. The explanation of both a higher methylation rate and its optimum at pH 6 suggests the activation of methylation by means of sulphhydryl-groups.
In vivo and in vitro phosphorylation of DNA-dependent RNA polymerase of Escherichia coli by bacteriophage-T7-induced protein kinase.
