Summary
We have analyzed some chemical properties of the sigma subunit of RNA polymerase from the sigma mutants: rpoD1 (Gross et al., 1978), rpoD2 (formerly known as alt-1) (Silverstone et al., 1972; Travers et al., 1978), and rpoD800 (Gross et al., 1979). Each of the three mutants is located at about 66 min on the E. coli genetic map and exhibits an alteration in the enzymatic properties of its sigma subunit. The tryptic peptides and isoelectric focusing behavior were analyzed for mutant and wild type sigma. A single, but different altered lysine tryptic peptide was observed for each mutant. No altered arginine tryptic peptides were observed. The rpoD800 mutant sigma showed an altered isoelectric point. These studies provide chemical evidence that the sigma polypeptide in all three mutants is altered and strongly support the conclusion that the mutations are in the structural gene for sigma.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachman, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of E. coli K-12. Bacteriol. Rev. 40, 116–167 (1976)
Burgess, R.R.: Separation and characterization of the subunits of RNA polymerase. J. Biol. Chem. 244, 6168–6176 (1969)
Burgess, R.R., Gross, C., Engback, F.: Cysteine and methionine content of E. coli RNA polymerase subunits. J. Bacteriol. 128, 390–393 (1976)
Burgess, R.R., Jendrisak, J.J.: A procedure for the rapid, large-scale purification of E. coli DNA-dependent RNA polymerase involving polymin P precipitation and DNA-cellulose chromatography. Biochemistry 14, 4634–4638 (1975)
Burgess, R.R., Travers, A.A., Dunn, J.J., Bautz, E.K.F.: Factor stimulating transcription by RNA polymerase. Nature 221, 43–46 (1969)
Fujiki, H., Palm, P., Zillig, W., Calendar, R., Sunshine, M.: Identification of a mutation within the structural gene of the alpha subunit of the RNA polymerase of E. coli. Mol. Gen. Genet. 145, 19–22 (1976)
Gross, C., Engbaek, F., Flammang, T., Burgess, R.: Rapid micromethod for the purification of E. coli RNA polymerase and the preparation of bacterial extracts active in RNA synthesis. J. Bacteriol. 128, 382–389 (1976)
Gross, C., Hoffman, J., Ward, C., Hager, D., Burdick, G., Berger, H. Burgess, R.: Mutation affecting thermostability of sigma subunit of E. coli RNA polymerase lies near the dnaG locus at about 66 mm on the E. coli gentic map. Proc. Natl. Acad. Sci. U.S.A. 75, 427–431 (1978)
Gross, C., Liebke, H., Walter, W., Burgess, R.: Function of the sigma subunit of E. coli RNA polymerase. I. Genetic Characterization of the mutant rpoD800 (submitted for publication)
Harris, J.D., Heilig, J.S., Martinez, I.I., Calendar, R., Isaksson, L.A.: A temperature-sensitive E. coli mutant affecting the sigma subunit of DNA-dependent RNA polymerase. Proc. Natl. Acad. Sci. U.S.A. 75, 6177–6181 (1978)
Harris, J.D., Martinez, I.I., Calendar, R.: A gene from E. coli affecting the sigma subunit of RNA polymerase. Proc. Natl. Acad. Sci. U.S.A. 74, 1836–1840 (1977)
Hirs, C.H.W.: Oxidation of ribonuclease with performic acid. J. Biol. Chem. 219, 611–621 (1956)
Jacobsen, M.F., Asso, J., and Baltimore, D.: Further evidence on the formation of poliovirus proteins. J. Mol. Biol. 49, 657–669 (1970)
Jaskunas, S.R., Burgess, R.R., Lindahl, L., Nomura, M.: Two clusters of genes for RNA polymerase and ribosome components in E. coli. In: RNA Polymerase (R. Losick, M. Chamberlin, eds.), pp. 539–552. New York: Cold Spring Harbor Laboratory 1976
Jaskunas, S.R., Burgess, R.R., Nomura, M.: Identification of a gene for the alpha subunit of RNA polymerase at the str-spc region of the E. coli chromosome. Proc. Natl. Acid. Sci. U.S.A. 72, 5036–5040 (1975)
Laemmli, U.K. Cleavage of structural proteins during the assembly of the head of phage T4. Nature 237, 680–685 (1970)
Lowe, P.A., Hager, D.A., Burgess, R.R.: Purification and properties of the sigma subunit of E. coli DNA-dependent RNA polymerase. Biochemistry 18, 1344–1352 (1979)
Mosser, A.G., Montelaro, R.C., and Rueckert, R.R.: Proteins of Rousassociated virus type 61: polypeptide stoichiometry and evidence that gp35 is not a cleavage product of a gp85. J. Virol. 23, 10–19 (1977)
Nakamura, Y.: RNA polymerase mutant with altered sigma factor in E. coli. Mol. Gen. Genet. 105, 1–6 (1978)
Nakamura, Y., Osawa, T., Yura, T.: Chromosomal location of a structural gene for the RNA polymerase sigma factor in E. coli. Proc. Natl. Acad. Sci. U.S.A. 74, 1831–1835 (1977)
Portalier, R.C., Robert-Baudouy, J.M., Nemoz, G.M.: Studies of mutations in the uronic isomerase and altronic oxidoreductase structural genes of E. coli K12. Mol. Gen. Gent. 128, 301–319 (1974)
Scaife, J.: Bacterial RNA polymerases: The genetics and control of their synthesis. In: RNA polymerase (R. Losick and M. Chamberlin, eds.), pp. 207–225. New York: Cold Spring Harbor Laboratory 1976
Silverstone, A.I., Goman, M., Scaife, J.G.: ALT: A new factor involved in the synthesis of RNA by E. coli. Mol. Gen. Genet. 118, 223–234 (1972)
Travers, A.A., Buckland, R., Goman, M., LeGrice, S.S.G., Scaife, J.G.: A mutation affecting the sigma subunit of RNA polymerase changes transcriptional specificity. Nature 273, 354–358 (1978)
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Burgess, R.R., Gross, C.A., Walter, W. et al. Altered chemical properties in three mutants of E. coli RNA polymerase sigma subunit. Molec. gen. Genet. 175, 251–257 (1979). https://doi.org/10.1007/BF00397224
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00397224