Skip to main content
Log in

A new mutation rpoD800, affecting the sigma subunit of E. coli RNA polymerase is allelic to two other sigma mutants

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

We have characterized a new mutation rpoD800 affecting the sigma gene of E. coli. Upon transfer to high temperature, a strain with the rpoD800 mutation ceases growth within 30 min. We find that this mutation renders sigma about 10-fold more thermolabile than the wild type sigma at 45°C in vitro. We have compared the temperature profile for inactivation of wild type and mutant sigma and find that the mutant inactivates at a temperature about 9° C lower than does the wild type.

The chromosomal locus affected by rpoD800 is shown to be allelic to the locus affected by the spontaneous mutants ts285 and alt-1. All three mutations result in altered sigma and in altered growth at high temperature. We argue that the single locus affected is the structural gene for the sigma subunit of E. coli RNA polymerase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Bacteriol. Rev. 40, 116–167 (1976)

    Google Scholar 

  • Bonhoeffer, F.: DNA transfer and DNA synthesis during bacterial conjugation. Z. Vererbungsl. 98, 141–149 (1966)

    Google Scholar 

  • Burgess, R.R., Gross, C.A., Walter, W., and Lowe, P.A.: Altered chemical properties in three mutants of E. coli RNA polymerase sigma subunit. Mol. Gen. Genet. 175, 251–257 (1979)

    Google Scholar 

  • 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)

    Google Scholar 

  • Burgess, R.R., Travers, A.A., Dunn, J.J., Bautz, E.K.F.: Factor stimulating transcription by RNA polymerase. Nature 221, 43–46 (1969)

    Google Scholar 

  • Chen, P.L., Carl, P.L. Genetic map location of the E. coli dnaG gene. J. Bacteriol 124, 1613–1614 (1975)

    Google Scholar 

  • Gross, C., Engbaek, F., Flammang, T., Burgess, R.: Rapid micro-method for the purification of E. coli RNA polymerase and the preparation of bacterial extracts active in RNA synthesis. J. Bacteriol. 128, 382–389 (1976)

    Google Scholar 

  • 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 min on the E. coli genetic map. Proc. Natl. Acad. Sci. U.S.A. 75, 427–431 (1978)

    Google Scholar 

  • Gross, J.: Curr. Top. Microbiol. Immunol. 57, 39 (1972)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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)

    Google Scholar 

  • Lark, K.G.: Evidence for the direct involvement of RNA in the initiation of DNA replication in E. coli 15T- J. Mol. Biol. 64, 47–60 (1972)

    Google Scholar 

  • 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)

    Google Scholar 

  • Miller, J.: Experiments in molecular genetics Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory 1972

    Google Scholar 

  • Nakamura, Y.: RNA polymerase mutant with altered sigma factor in E. coli. Mol. Gen. Genet. 105, 1–6 (1978)

    Google Scholar 

  • 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)

    Google Scholar 

  • 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. Genet. 128, 301–319 (1974)

    Google Scholar 

  • 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)

    Google Scholar 

  • Travers A.A., Buckland, R., Goman, M., LeGrice, S.S.G., Scaife, J.G.: A mutation affecting the sigma subunit of RNA polymerase change transcriptional specificity. Nature 273, 354–358 (1978)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liebke, H., Gross, C., Walter, W. et al. A new mutation rpoD800, affecting the sigma subunit of E. coli RNA polymerase is allelic to two other sigma mutants. Molec. Gen. Genet. 177, 277–282 (1980). https://doi.org/10.1007/BF00267439

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00267439

Keywords

Navigation