Summary
The assay for the ksgA-encoded S-adenosylmethionine-— 6-N′, N′-adenosyl (rRNA) dimethyltransferase has been improved; the gel-filtration molecular weight of partially purified enzyme under two different sets of conditions was found to be 55,000 or 26,000 daltons. We have determined methyltransferase activities in strains where ksgA was brought under the control of the mitomycin C-inducible promoter of the colicin El gene. Our studies show that ksgA is transcribed counterclockwise on the Escherichia coli chromosome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andrésson, Ó.S., Davies, J.E.: Lambda transducing phages for the E. coli genes ksgA and pdxA. Mol. Gen. Genet. 179, 201–209 (1980a)
Andrésson, O.S., Davies, J.E.: Genetic organization and restriction enzyme cleavage map of the ksgA-pdxA region of the Escherichia coli chromosome. Mol. Gen. Genet. 179, 211–216 (1980b)
Barnes, W.M.: Plasmid detection and sizing in single colony lysates. Science 195, 393–394 (1977)
Bernard, H.-U., Helinski, D.R.: Use of the phage lambda promoter P L to promote gene expression in hybrid plasmid cloning vehicles. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan, eds.), in press. New York: Academic Press 1979
Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976)
Davies, J.E., Smith, D.: Plasmid-determined resistance to antimicrobial agents. Annu. Rev. Microbiol. 32, 469–518 (1978)
Haas, M.J., Dowding, J.E.: Aminoglycoside-modifying enzymes. In: Methods in enzymology (J.H. Hash, ed.), Vol XLIII, pp. 611–628. New York: Academic Press 1975
Helser, T.L., Davies, J.E., Dahlberg, J.E.: Mechanism of kasugamycin resistance in Escherichia coli. Nature New Biol. 235, 6–9 (1972)
Klein, R.D., Selsing, E., Wells, R.D.: A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid, in press (1980)
Lennox, E.S.: Translation of linked genetic characters of the host by bacteriophage P1. Virology 1, 190–206 (1980)
Miller, J.H.: Experiments in molecular genetics. New York: Cold Spring Harbor Laboratory 1972
Murphey, W.H., Kitto, G.B., Everse, J., Kaplan, N.O.: Malate dehydrogenases. I. A survey of molecular size measured by gel filtration. Biochemistry 6, 603–610 (1967)
Ochoa, S.: Crystalline condensing enzyme from pig heart. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan, eds.) Vol. I, pp. 685–694. New York: Academic Press 1955a
Ochoa, S.: Malic dehydrogenase from pig heart. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan, eds.) Vol. I, pp. 735–739. New York: Academic Press 1955b
Patient, R.K.: Characterization of in vitro transcription and termination sites in ColEl DNA. Nucleic Acids 6, 2647–2665 (1979)
Poldermans, B., Roza, L., Van Knippenberg, P.H.: Studies on the function of two adjacent N6,N6-dimethyladenosines near the 3'end of 16S ribosomal RNA of Escherichia coli. III. Purification and properties of the methylating enzyme and methylase-30S interactions. J. Biol. Chem. 254, 9094–9100 (1979)
Postle, K., Reznikoff, W.S.: HindII and HindIII restriction maps of the attϕ80-tonB-trp region of the Escherichia coli genome, and location of the tonB gene. J. Bacteriol. 136, 1165–1173 (1978)
Selker, E., Brown, K., Yanofsky, C.: Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColEl. J. Bacteriol. 129, 388–394 (1977)
So, M., Gill, R., Falkow, S.: The generation of a ColEl-Apr cloning vehicle which allows detection of inserted DNA. Mol. Gen. Genet. 142, 239–249 (1975)
Tabor, C.W.: Adenosylmethionine decarboxylase. In: Methods in enzymology (S.P. Colowick and N.O. Kaplan eds.), Vol. V, pp. 756–760. New York: Academic Press 1962
Tanaka, T., Weisblum, B.: Construction of a colicin El-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J. Bacteriol. 121, 354–362 (1975)
Thammana, P.: Studies on the anatomy of ribosomes from Escherichia coli. Ph.D. dissertation, University of Wisconsin-Madison, 1974
Thammana, P., Cantor, C.R.: Studies on ribosome structure and interactions near the m 62 A m 62 A sequence. Nucleic Acids 5, 805–823 (1978)
Thammana, P., Held, W.A.: Methylation of 16S RNA during ribosome asembly in vitro. Nature 251, 682–686 (1974)
Traub, P., Mizushima, S., Lowry, C.V., Nomura, M.: Reconstitution of ribosomes from subribosomal components. In: Methods in enzymology (K. Moldave and L. Grossman, eds.), Vol. XX, pp. 391–407, New York: Academic Press 1971
Wu, J.-Y., Yang, J.T.: Physicochemical characterization of citrate synthase and its subunits. J. Biol. Chem. 245, 212–218 (1970)
Author information
Authors and Affiliations
Additional information
Communicated by K. Isono
Rights and permissions
About this article
Cite this article
Andrésson, Ó.S., Davies, J.E. Some properties of the ribosomal RNA methyltransferase encoded by ksgA and the polarity of ksgA transcription. Molec. Gen. Genet. 179, 217–222 (1980). https://doi.org/10.1007/BF00268466
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268466