Summary
Crude extracts of bacteria lysogenic for temperate phage Mu contain proteins that retain specifically Mu DNA on nitrocellulose filters. The amount of binding protein is directly proportional to the number of Mu prophages per E. coli genome. Specificity of the binding reaction could be demonstrated by using heterologous DNAs as substrate and by a competition experiment. By using hybrid plasmids containing different amounts of the immunity end and extending to various degrees into Mu DNA, it was found that the binding activity is coded for by the left 1,000 nucleotide-pair HindIII fragment. When using these hybrid plasmids as binding substrate, two different binding sites for the immunity product were detected. Joining of the MucI gene to the left λ early promoter resulted in increased production of immunity protein at elevated temperature. A possible explanation for the relatively low amounts of immunity protein in all of the different strains studied is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abelson, J., Boram, W., Bukhari, A.I., Faelen, M., Howe, M.M., Metlay, M., Taylor, A.K., Toussaint, A., Putte, P. van de: Summary of the genetic mapping of prophage Mu. Virology 54, 90–92 (1973)
Allet, B., Bukhari, A.I.: Analysis of bacteriophage Mu and lambda-Mu hybrid DNA by specific endonucleases. J. Mol. Biol. 92, 529–540 (1975)
Bade, E.G.: Asymmetric transcription of bacteriophage Mu-1. J. Virol. 10, 1205–1207 (1972)
Ballivet, M., Reichardt, L.F., Eisen, H.: Purification properties of coliphage 21 repressor. Eur. J. Biochem. 73, 601–606 (1977)
Bezdek, A., Amati, P.: Properties of P22 and related Salmonella phage. I. General features and host specificity. Virology 31, 272–278 (1967)
Bolivar, F., Rodriguez, R.L., Betlach, M.C., Boyer, H.W.: Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene 2, 75–93 (1977a)
Bolivar, F., Rodriguez, R.L., Greene, P.J., Betlach, M.C., Heyneker, H.K., Boyer, H.W.: Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2, 95–113 (1977b)
Bolivar, F.G.: Modificación de la estructura del DNA bacteriano por un mutàgeno biológico. Ph. D. Thesis, Universidad National Autónoma, Mexico (1971)
Botstein, D.: Synthesis and maturation of phage P22 DNA. I. Identification of intermediates. J. Mol. Biol. 34, 621–641 (1968)
Bukhari, A.I., Zipser, D.: Random insertion of Mu-1 DNA within a single gene. Nature New Biol. 236, 240–243 (1972)
Echols, H., Green, L.: Establishment and maintenance of repression by bacteriophage lambda: The role of the cI, cII, and cIII proteins. Proc. Natl. Acad. Sci. U.S.A. 68, 2190–2194 (1971)
Faelen, M., Toussaint, A., Lafonteyne, J. de: Model for the enhancement of λ-gal integration into partially induced Mu-1 lysogens. J. Bacteriol. 121, 873–882 (1975)
Figurski, D., Meyer, R., Miller, D., Helinski, R.: Generation in vitro of deletions in the broad host range plasmid RK2 using phage Mu insertions and a restriction endonuclease. Gene 1, 107–119 (1976)
Hedgepeth, J., Ballivet, M., Eisen, H.: Lambda phage promoter used to enhance expression of a plasmid-cloned gene. Mol. Gen. Genet. 163, 197–203 (1978)
Herskowitz, I.: Control of gene expression in bacteriophage lambda. Annu. Rev. Genet. 7, 289–324 (1973)
Howe, M.M.: Genetic studies on bacteriophage Mu. Ph.D. Thesis, Massachusetts Institude of Technology (1972)
Howe, M.M., Bade, E.G.: Molecular biology of bacteriophage Mu. Science 190, 624–632 (1975)
Lowry, O.H., Rosebrough, N.K., Farr, A.K., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Riggs, A.D., Bourgeois, S., Newby, R.F., Cohn, M.: DNA binding of the lac repressor. J. Mol. Biol. 34, 365–368 (1968)
Rodriguez, R.K., Bolivar, F., Goodman, H.M., Boyer, H.W., Betlach, M.: Construction and characterization of cloning vehicles. In: Molecular mechanisms in the control of gene expression (Nierlich, D.P., Rutter, W.J., Fox, C.F., eds.), Vol. 5, pp. 471–477. ICN-UCLA Symposia on Molecular and Cellular Biology. New York: Academic Press 1976
Schumann, W., Bade, E.G.: Bacteriophage-specific DNA-binding proteins in P22-lysogenic and in P22-infected Salmonella typhimurium. J. Virol. 20, 334–338 (1976)
Schumann, W., Bade, W.G.: In vivo constructed plasmids containing both ends of bacteriophage Mu DNA express phage functions. Mol. Gen. Genet. 169, 97–105 (1979)
Schumann, W., Bade, E.G., Delius, H., Hübert, P.: Cloning of a restriction fragment of phage Mu DNA coding for early functions. Mol. Gen. Genet. 160, 115–118 (1978)
So, M., Gill, R., Falkow, S.: The generation of a colE1-ApR cloning vehicle which allows detection of inserted DNA. Mol. Gen. Genet. 124, 239–249 (1975)
Susskind, M.M., Botstein, D.: Molecular genetics of bacteriophage P22. Microbiol. Reviews 42, 385–413 (1978)
Taylor, A.L.: Bacteriophage induced mutation in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 50, 1043–1051 (1963)
Wijffelman, C., Lotterman, B.: Kinetics of Mu DNA synthesis. Mol. Gen. Genet. 151, 169–174 (1977)
Wu, A.M., Gosh, S., Echols, H., Spiegelman, W.G.: Repression by the cI protein of phage λ: in vitro inhibition of RNA synthesis. J. Mol. Biol. 67, 407–421 (1972)
Zipser, D., Moses, P., Kahmann, R., Kamp, D.: The molecular cloning of the immunity gene of phage Mu. Gene 2, 263–271 (1977)
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
This work was supported by the Deutsche Forschungsgemeinschaft (grant Ba 600/1)
Rights and permissions
About this article
Cite this article
Schumann, W., Westphal, C., Bade, E.G. et al. Origin and binding specificity of protein(s) coded for by Mu prophages. Molec. gen. Genet. 173, 189–196 (1979). https://doi.org/10.1007/BF00330310
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00330310