Summary
A fragment of Escherichia coli bacteriophage T4D DNA, containing 6.1 Kbp which included the six genes (genes 25, 26, 51, 27, 28 and 29) coding for the tail baseplate central plug has been partially characterized. This DNA fragment was obtained originally by Wilson et al. (1977) by the action of the restriction enzyme EcoRI on a modified form of T4 DNA and was inserted in the pBR322 plasmid and then incorporated into an E. coli K12 strain called RRI. This plasmid containing the phage DNA fragment has now been reisolated and screened for cleavage sites for various restriction endonucleases. Restriction enzymes Bgl 11 and Xbal each attacked one restriction site and the enzyme Hpa 1 attacked two restriction sites on this fragment. The combined digestion of the hybrid plasmid containing the T4 EcoRI DNA fragment conjugated to the pBR322 plasmid with one of these enzymes plus Bam H1 restriction enzyme resulted in the localization of the restriction site for Bgl 11, Xba 1 and Hpa 1.
Escherichia coli strain B cells were transformed with this hybrid plasmid and found to have some unexpected properties. E. coli B cells, which are normally restrictive for T4 amber mutants and for T4 temperature sensitive mutants (at 44°) after transformation, were permissive for 25am, 26am and 26Ts, 51am, and 51Ts, 27Ts, and 28Ts T4 mutants. Extracts from the transformed E. coli cells were found in complementation experiments to contain the gene 29 product, as well as the gene 26 product, the gene 51 product, and the gene 27 product. The complementation experiments and the permissiveness of the transformed E. coli B cells to the various conditional lethal mutants clearly showed that the six T4 genes were producing all six gene products in these transformed cells. However, these cells were not permissive for T4 amber mutants in genes 27, 28, and 29. The transformed E. coli B cells, as compared to untransformed cells, were found to have altered outer cell walls which made them highly labile to osmotic shock and to an increased rate of killing by wild type T4 and all T4 amber mutants except for T4 am29. The change in cell walls of the transformed cells has been found to be due to the T4 baseplate genes on the hybrid plasmid, since E. coli B transformed by the pBR322 plasmid alone does not show the increase in osmotic sensitivity.
Similar content being viewed by others
References
Adams MJ (1959) Bacteriophages. Interscience Publishers Inc, New York
Edgar RS, Wood WB (1966) Morphogenesis of bacteriophage T4 in extracts of mutant-infected cells. Proc Natl Acad Sci USA 55:498–505
Kikuchi Y, King J (1975a) Genetic control of bacteriophage T4 baseplate morphogenesis. II. Mutants unable to form the central part of the baseplate. J Mol Biol 99:673–694
Kikuchi Y, King J (1975b) Genetic control of bacteriophage T4 baseplate morphogenesis. III. Formation of the central plug and overall assembly pathway. J Mol Biol 99:695–716
Kozloff LM, Zorzopulos J (1981) Dual functions of bacteriophage T4D gene of product: Structural component of the viral tail baseplate central plug and cleavage enzyme for folyl polyglutamates. I. Identification of T4D gene 28 product in the tail plug. J Virol 40:635–644
Kutter E, Beng A, Sluss R, Jensen L, Bradley D (1975) The production of undegraded cytosine-containing DNA by bacteriophage T4 in the absence of cDTPase and endonucleases II and IV, and its effects on T4-directed protein synthesis. J Mol Biol 99:591–607
Mattson T, Houwe G, van Bolle A, Selzer G, Epstein R (1977) Genetic identification of cloned fragment of bacteriophage T4 DNA and complementation by some clones containing early genes. Mol Gen Genet 154:319–326
Neidhardt FC, Wirth R, Smith MW, Bogelen RV (1980) Selective synthesis of plasmid-coded proteins by Escherichia coli during recovery from chloramphenicol treatment. J Bacteriol 143:535–537
Ratner D (1974) Bacteriophage T4 transcriptional control. Gene 55 codes for a protein bound to Escherichia coli RNA polymerase. J Mol Biol 89:803–807
Snyder LR, Montgomery DL (1974) Inhibition of T4 growth by an RNA polymerase mutation of Escherichia coli physiological and genetic analysis of the effects during phage development. Virology 62:184–196
Stevens A (1972) New small polypeptides associated with DNA-dependent RNA polymerase of Escherichia coli after infection with bacteriophage T4. Proc Natl Acad Sci USA 69:603–607
Velten J, Fukada K, Abelson J (1976) In vitro construction of bacteriophage λ and plasmid DNA molecules containing DNA fragments from bacteriophage T4. Gene 1:93–1061
Wilson GG, Tanyashin VI, Murray NE (1977) Molecular cloning of fragments of bacteriophage T4 DNA. Mol Gen Genet 156:203–214
Wood WB, Revel HR (1976) The genome of bacteriophage T4. Bacteriol Rev 40:847–868
Yee JK, Marsh RC (1981) Alignment of a Restriction Map with the Genetic map of bacteriophage T4. J Virol 38:115–124
Zorzopulos J, Kozloff LM, Chapman V, DeLong S (1979) Bacteriophage T4D receptors and the Escherichia coli cell wall structure: Role of spherical particles and protein b of the cell wall in bacteriophage infection. J Bacteriol 137:545–555
Author information
Authors and Affiliations
Additional information
Communicated by H.W. Boyer
Rights and permissions
About this article
Cite this article
Zorzopulos, J., Kozloff, L.M. A T4 DNA fragment containing genes for the baseplate central plug: Endonuclease restriction, gene expression and cell wall changes. Molec Gen Genet 187, 278–285 (1982). https://doi.org/10.1007/BF00331129
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331129