Summary
Four mutants of Escherichia coli KL16 resistant to the antibiotic Thiolutin have been isolated. This drug was earlier reported to be an inhibitor of RNA chain elongation. The first mutant, TLrI, is resistant only in rich or partially rich media: it can, however, grow in minimal medium containing the drug with a very long doubling time. The other mutants TLrII, TLrIIIa and TLrIIIb are resistant in rich as well as minimal media. β-galactosidase could not be induced in TLrI and TLrII in the presence of thiolutin whereas the enzyme is constitutively synthesised in TLrIIIa and TLrIIIb irrespective of the drug.
The mutants do not support the development of phage T4 in presence of the drug, if the drug is added along with the phage, but “escape” the inhibition if phage development is allowed to proceed for some time before the addition of the drug. The time of this escape is characteristic of the mutant. Even in a sensitive strain, T7 growth escapes inhibition very soon after infection, around the time the phage-specific RNA polymerase is synthesised. In the parent strain the kinetics of inhibition of β-galactosidase induction resembles more the inhibition caused by rifampicin than by streptolydigin. It is proposed that thiolutin could be an inhibitor of RNA chain initiation and resistance might be due to mutation in the subunit(s)/factor(s) involved in initiation.
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References
Adelberg, E.A., Mandel, M., Chen, G.C.C.: Optimal conditions for mutagenesis by N-methyl, N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem. biophys. Res. Commun. 18, 788–795 (1965)
Bremer, H., Yuan, D.: Chain growth rate of messenger RNA in Escherichia coli infected with bacteriophage T4. J. molec. Biol. 34, 527–540 (1968)
Cassani, G., Burgess, R.R., Goodman, H.: Streptolydigin inhibition of RNA polymerase. Cold Spr. Harb. Symp. quant. Biol. 35, 59–63 (1970)
Celmer, W.D., Solomon, I.A.: The structures of thiolutin, aureothricin, antibiotics containing a unique pyrrolinodithiol nucleus. J. Amer. chem. Soc. 77, 2861 (1955)
Chakrabarti, S.L., Maitra, P.K.: Development of auxotrophy by streptomycin resistant mutation. J. Bact. 118, 1179–1180 (1974)
Chamberlin, M., McGrath, J.: Characterization of a T7 specific RNA polymerase isolated from E. coli infected with T7 phage. Cold Spr. Harb. Symp. quant. Biol. 35, 259–262 (1970)
Errington, L., Glass, R.E., Hayward, R.S., Scaife, J.G.: Structure and orientation of an RNA polymerase operon in Escherichia coli. Nature (Lond.) 249, 519–522 (1974)
Haselkorn, R., Vogel, M., Brown, R.D.: Conservation of the rifamicyn sensitivity of transcription during T4 development. Nature (Lond.) 221, 836–838 (1969)
Heil, A., Zillig, W.: Reconstitution of bacterial DNA-dependent RNA polymerase from isolated subunits as a tool for the elucidation of the role of the subunits in transcription. FEBS Letters 11, 165–168 (1970)
Iwakura, Y., Ishihama, A., Yura, J.: RNA polymerase mutants of E. coli. Molec. gen. Genet. 121, 181–196 (1973)
Jayaraman, R.: Transcription of bacteriophage T4 DNA by Escherichia coli RNA polymerase in vitro: Identification of some immediate early and delayed early genes. J. molec. Biol. 70, 253–263 (1972)
Jimenz, A., Tipper, D.J., Davies, J.: Mode of action of thiolutin, an inhibitor of macromolecular synthesis in Saccharomyces cerevisiae. Antimicrob. Ag. Chemother. 3, 729–738 (1973)
Khachatourians, G.G., Tipper, D.J.: Inhibition of messenger Ribonucleic acid synthesis in Escherichia coli by thiolutin. J. Bact. 119, 795–804 (1974a)
Khachatourians, G.G., Tipper, D.J.: In vivo effect of Thiolutin on cell growth and macromolecular synthesis in Escherichia coli. Antimicrob. Ag. Chemother 6, 304–310 (1974b)
Leive, L.: Actinomycin sensitivity in Escherichia coli produced by EDTA. Biochem. biophys. Res. Commun. 18, 13–17 (1965)
Lill, H., Lill, U., Sippel, A., Hartmann, G.: The inhibition of the RNA polymerase reaction by rifampicin. Lepetit. colloq. on RNA polymerase and Transcription (L. Silvestri ed.), p. 55–64. Amsterdam: North-Holland Publ. Co. 1969
Mäkelä, O., Mäkelä, P.H., Soikkeli, S.: Sex specificity of the bacteriophage T7. Ann. Med. exp. Fenn. 42, 188–195 (1964)
Mauro, E. di, Snyder, L., Marino, P., Lamberti, A., Coppo, A., Tocchini-Valentini: Ritampicin sensitivity of the components of DNA dependent RNA polymerase. Nature (Lond.) 222, 533–538 (1969)
Miller, J.H.: Experiments in molecular genetics. Cold Spring Harbor, N.Y.: Cold spring Harbor Laboratory 1972
Pitale, M.P., Jayaraman, R.: Transcription of T4 genome in vitro: Heterogeneity of RNA polymerase in crude extracts of normal and T4 infected Escherichia coli B. Biochemistry 14, 1265–1271 (1975)
Rabussay, D., Zillig, W.: A rifampicin resistant RNA polymerase from E. coli altered in the β subunit. FEBS Letters 5, 104–105 (1969)
Schleif, R.: Isolation and characterisation of a streptolydigin resistant RNA polymerase. Nature (Lond.) 223, 1068–1069 (1969)
Siddhikol, C., Erbstoeszer, J.W., Weisblum, B.: Mode of action of Streptolydigin. J. Bact. 99, 151–155 (1969)
Tipper, D.J.: Inhibition of yeast Ribonucleic acid polymerase by thiolutin. J. Bact. 116, 245–256 (1973)
Tocchini-Valentini, G.P., Marino, P., Colvill, A.J.: Mutant of E. coli containing an altered DNA depedent RNA polymerase. Nature (Lond.) 220, 275–276 (1968)
Travers, A.A.: RNA polymerase and T4 development. Cold Spr. Harb. Symp. quant. Biol. 35, 241–251 (1970)
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Sivasubramanian, N., Jayaraman, R. Thiolutin resistant mutants of Escherichia coli are they RNA chain initiation mutants?. Molec. gen. Genet. 145, 89–96 (1976). https://doi.org/10.1007/BF00331562
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DOI: https://doi.org/10.1007/BF00331562