Summary
Host controlled restriction in Escherichia coli can be relieved by pre-infecting restricting cells with modified λ helper phages. This process, in which intact unmodified phage genomes are allowed to escape restriction attack, is mediated by a newly identified λ function called ral. The ral gene has been located by deletion mapping between cIII and N. Efficient expression of the ral gene requires the product of the regulator gene N. Polyacrylamide gel analysis of the λ proteins specified by the cIII-N region failed to reveal the product of the ral gene, but demonstrated that protein Ea10 is encoded by a gene located immediately to the left of ral. From these results the map order cIII-Ea10-ral-T L1-N was deduded.
Ral specifically alleviates restriction in E. coli K and E. coli B, but does not affect restriction systems EcoRI, EcoRII and EcoP1. In addition, ral enhances the modification activity of the EcoK and EcoB restriction enzymes: we observed that efficient modification of progeny phages obtained by propagating unmodified λ phages in r- m+ hosts, is dependent upon the presence of ral. We thus conclude that the ral gene product acts by modulating the restriction and modification activities of the type I restriction systems in E. coli, and the possible mechanisms will be discussed.
Similar content being viewed by others
References
Adhya, S., Gottesman, M., De Crombrugghe, B.: Release of polarity in Escherichia coli by gene N of phage lambda: termination and antitermination of transcription. Proc. Natl. Acad. Sci. U.S.A. 71, 2534–2538 (1974)
Appleyard, R.K.: Segregation of new lysogenic types during growth of a double lysogenic strain derived from Escherichia coli K12. Genetics 39, 440–452 (1954)
Arber, W.: DNA modification and restriction. Prog. Nucleic Acid Res. Mol. Biol. 14, 1–37 (1974)
Arber, W., Lataste-Dorolle, C.: Erweiterung des Wirtsbereiches des Bacteriophage Lambda auf Escherichia coli B. Pathol. Microbiol. (Basel) 24, 1012–1018 (1961)
Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K12. Bacteriol. Rev. 40, 116–166 (1976)
Bertani, G.: Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J. Bacteriol. 62, 293–300 (1951)
Bertani, G., Weigle, J.J.: Host controlled variation in bacterial viruses. J. Bacteriol. 65, 113–121 (1953)
Boyer, H.W.: DNA restriction and modification mechanisms in bacteria. Annu. Rev. Microbiol. 25, 135–176 (1971)
Boyer, H.W.: Roulland-Dussoix, D.: A complementation analysis of restriction and modification in Escherichia coli. J. Mol. Biol. 41, 459–472 (1969)
Castellazzi, M., Brachet, P., Eisen, H.: Isolation and characterization of deletions in bacteriophage lambda residing as prophage in E. coli K12. Mol. Gen. Genet. 117, 211–218 (1972)
Colson, C., Glover, S.W., Symonds, N., Stacey, K.A.: The location of the genes for host controlled modification and restriction in Escherichia coli K12. Genetics 52, 1043–1050 (1965)
Davidson, N., Szybalski, W.: Physical and chemical characteristics of lambda DNA. In: The bacteriophage lambda (A.D. Hershey, ed.), pp. 45–82. New York: Cold Spring Harbor Laboratory 1971
Debrouwere, L., Van Montagu, M., Schell, J.: The ral gene of phage λ. III. Interference with E. coli ATP dependent functions. Mol. Gen. Genet. 179, 81–88 (1980b)
Debrouwere, L., Zabeau, M., Van Montagu, M., Schell, J.: The ral gene of phage λ. II. Isolation and characterization of ral dificient mutants. Mol. Gen. Genet. 179, 75–80 (1980a)
Dussoix, D., Arber, W.: Host specificity of DNA produced by Escherichia coli. II. Control and acceptance of DNA from infeeting phage lambda. J. Mol. Biol. 5, 37–49 (1962)
Eskin, B., Linn, S.: The deoxyribonucleic acid modification and restriction enzymes from Escherichia coli B. II. Purification, subunit structure and catalytic properties of the restriction endonuclease. J. Biol. Chem. 274, 6183–6191 (1972)
Franklin, N.C.: Altered reading of genetic signals fused to the N operon of bacteriophage lambda: genetic evidence for modification of polymerase by the protein product of the N gene. J. Mol. Biol. 89, 33–48 (1974)
Gottesman, M.M., Gottesman, M.E., Gottesman, S., Gellert, M.: Characterization of lambda reverse as an E. coli phage carrying a unique set of host-derived recombination functions. J. Mol. Biol. 88, 471–487 (1974)
Greer, H.: The kil gene of bacteriophage lambda. Virology 66, 589–604 (1975)
Haberman, A., Heywood, J., Meselson, M.: DNA modification methylase activity of Escherichia coli restriction endonucleases K and P. Proc. Natl. Acad. Sci. U.S.A. 69, 3138–3141 (1972)
Heip, J., Rolfe, B., Schell, J.: Abolition of host cell restriction by high multiplicity of phage infection. Virology 59, 356–370 (1974)
Hendrix, R.: Identification of proteins coded in phage lambda. In: The bacteriophage lambda (A.D. Hershey, ed.), pp. 355–370. New York: Cold Spring Harbor Laboratory 1971
Hoekstra, W.P.M.: Restriction and modification of deoxyribonucleic acid in Escherichia coli. Ph. D. thesis, State University of Utrecht, Utrecht, The Netherlands (1966)
Hubacek, J., Glover, S.W.: Complementation analysis of temperature sensitive host specificity mutations in Escherichia coli. J. Mol. Biol. 50, 111–127 (1976)
Jacob, F., Wollman, E.L.: Etude génétique d'un bactériophage tempéré d'Escherichia coli. I. Le système génétique du bactériophage lambda. Ann. Inst. Pasteur Lille 87, 653–673 (1954)
Jacob, F., Wollman, E.L.: Sur les processus de conjugaison et de recombinaison chez Escherichia coli. I. L'induction par conjugaison ou induction zygotique. Ann. Inst. Pasteur Lille 91, 486–510 (1956)
Kaiser, A.D.: Mutations in a temperature bacteriophage affecting its ability to lysogenize Escherichia coli. Virology 3, 42–61 (1957)
Kayajanian, G.: Studies on the genetics of biotin-transducing, defective variants of bacteriophage lambda. Virology 36, 30–41 (1968)
Kellenberger, G., Zichichi, M.L., Weigle, J.: A mutation affecting the DNA content of bacteriophage lambda and its lysogenizing abilities. J. Mol. Biol. 3, 399–408 (1961)
Krüger, D.H., Schroeder, C., Hansen, S., Rosenthal, H.A.: Active protection by bacteriophages T3 and T7 against E. coli B and K specific restriction of their DNA. Mol. Gen. Genet. 153, 99–106 (1977)
Lomax, M.S., Greenberg, G.R.: Characteristics of the deo operon: Role in thymine utilization and sensitivity to deoxyribonucleosides. J. Bacteriol. 96, 501–514 (1968)
Manly, K.F., Signer, E.R., Radding, C.M.: Non-essential functions of bacteriophage lambda. Virology 37, 177–188 (1969)
Matsushiro, A.: Specialized transduction of tryptophan makers in Escherichia coli K12 by bacteriophage ϕ80. Virology 19, 475–482 (1963)
Meselson, M., Yuan, R., Heywood, J.: Restriction and modification of DNA. Annu. Rev. Biochem. 41, 447–466 (1971)
Miller, J.H.: Experiments in molecular genetics, p. 466. New York: Cold Spring Harbor Laboratory 1972
Pardee, A.B., Jacob, F., Monod, J.: The genetic control and cytoplasmic expression of inducibility in the synthesis by E. coli. J. Mol. Biol. 1, 165–178 (1959)
Parkinson, J.S., Huskey, R.J.: Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization. J. Mol. Biol. 56, 369–384 (1971)
Phashne, M.: Isolation of the lambda phage repressor. Proc. Natl. Acad. Sci. U.S.A. 57, 306–313 (1967)
Ratner, D.: Evidence that mutations in the SuA polarity suppressing gene directly affect termination factor rho. Nature 259, 151–153 (1976)
Roberts, J.W.: The rho factor: termination and antitermination in λ. Cold Spring Harbor Symp. Quant. Biol. 35, 121–126 (1970)
Salström, J.S., Szybalski, W.: Transcription termination sites in the major leftward operon of coliphage lambda. Virology 88, 256–262 (1978)
Schell, J., Glover, S.W.: The effect of heat on host controlled restriction of phage lambda in Escherichia coli K (P1). J. Gen. Microbiol. 45, 61–72 (1966)
Signer, E.R., Weil, J.: Recombination in bacteriophage lambda. I. Mutants deficient in general recombination. J. Mol. Biol. 34, 261–271 (1968)
Signer, E.R., Manly, K.F., Brunstetter, M.: Deletion mapping of the cIII-N region of bacteriophage lambda. Virology 39, 137–141 (1969)
Simmon, V.F., Lederberg, S.: Degradation of bacteriophage lambda deoxyribonucleic acid after restriction by Escherichia coli K12. J. Bacteriol. 112, 161–169 (1972)
Stratern, A., Herskowitz, I.: Defective prophage in Escherichia coli K12 strains. Virology 67, 136–143 (1975)
Studier, F.W.: Gene 0.3 of bacteriophage T7 acts to overcome the DNA restriction system of the host. J. Mol. Biol. 94, 283–295 (1975)
Studier, R., Movva, N.R.: SAMase gene of bacteriophage T3 is responsible for overcoming host restriction. J. Virol. 19, 136–145 (1976)
Sussman, R., Jacob, F.: Sur un système de répression thermosensible chez le bactériophage lambda d' Escherichia coli. Compt. Rend. Acad. Sci. Paris 254, 1517–1519 (1962)
Szybalski, W., Szybalski, E.H.: Physical mapping of the att-N region of coliphage lambda: apparent oversaturation of coding capacity in the gam-ral segment. Biochimie 56, 1497–1503 (1974)
Vovis, G.F., Horiuchi, K., Zinder, N.D.: Kinetics of methylation of DNA by a restriction endonuclease from Escherichia coli B. Proc. Natl. Acad. Sci. U.S.A. 71, 3810–3813 (1974)
Vovis, G.F., Zinder, N.D.: Methylation of f1 DNA by a restriction endonuclease from Escherichia coli B. J. Mol. Biol. 95, 557–568 (1975)
Wollman, E.L.: Transduction spécifique du marqueur biotine par le bactériophage lambda. Compt. Rend. Acad. Sci. Paris 257, 4225–4226 (1963)
Wood, W.B.: Host specificity of DNA produced by Escherichia coli: bacterial mutations affecting the restriction and modification of DNA. J. Mol. Biol. 16, 118–133 (1966)
Zabeau, M., Roberts, R.J.: The role of restriction endonucleases in molecular geneties. In: Molecular genetics (J.H. Taylor, ed.), Vol. III, pp. 1–69. New York: Academic Press 1979
Zabeau, M., Van Montagu, M., Schell, J.: The alleviation of host controlled restriction of unmodified phages by functions of bacteriophage lambda. Arch. Int. Physiol. Biochim. 81, 990 (1973)
Zissler, J., Signer, E.R., Schaefer, F.: The role of recombination in growth of bacteriophage lambda. II. Inhibition of growth by bacteriophage P2. In: The bacteriophage lambda (A.D. Hershey, ed.), pp. 469–475. New York Cold Spring Harbor Laboratory 1971
Author information
Authors and Affiliations
Additional information
Communicated by W. Arber
Rights and permissions
About this article
Cite this article
Zabeau, M., Friedman, S., Van Montagu, M. et al. The ral gene of phage λ. Molec. Gen. Genet. 179, 63–73 (1980). https://doi.org/10.1007/BF00268447
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268447