Rate of ribonucleic acid chain growth in Mycobacterium tuberculosis H37R(v)

Abstract

Two methods were employed to measure the rate of ribonucleic acid (RNA) chain growth in vivo in M. tuberculosis H37R(v) cultures, growing in Sauton medium at 37°C, with a generation time of 10 h. In the first, bacteria were allowed to assimilate [3H]uracil or [3H]guanine into their RNA for short time periods. The RNA was then extracted and hydrolyzed with alkali, and the radioactivity in the resulting nucleotides and nucleosides was measured. The data obtained by this method allowed the calculation of the individual nucleotide step times during the growth of RNA chains, from which the average rate of RNA chain elongation was estimated to be about 4 nucleotides per s. The second method employed the antibiotic rifampin, which specifically inhibits the initiation of RNA synthesis without interfering with the elongation and completion of nascent RNA chains. Using this method, the transcription time of the 16S, 23S, and 5S ribosomal RNA genes was estimated to be 7.6 min, which corresponds to a ribosomal RNA chain growth rate of 10 nucleotides per s.