Summary
Lysis protein T of phage T4 is required to allow the phage's lysozyme to reach the murein layer of the cell envelope and cause lysis. Using fusions of the cloned gene t with that of the Escherichia coli alkaline phosphatase or a fragment of the gene for the outer membrane protein OmpA, it was possible to identify T as an integral protein of the plasma membrane. The protein was present in the membrane as a homooligomer and was active at very low cellular concentrations. Expression of the cloned gene t was lethal without causing gross leakiness of the membrane. The functional equivalent of T in phage λ is protein S. An amber mutant of gene S can be complemented by gene t, although neither protein R of λ (the functional equivalent of T4 lysozyme) nor S possess any sequence similarity with their T4 counterparts. The murein-degrading enzymes (including that of phage P22) have in common a relatively small size (molecular masses of ca. 18 000) and a rather basic nature not exhibited by other E. coli cystosolic proteins. The results suggest that T acts as a pore that is specific for this type of enzyme.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altman E, Young K, Garrett J, Altman R, Young R (1985) Subcellular localization of lethal lysis proteins of bacteriophage λ and ϕX174. J Virol 53:1008–1011
Bienkowska-Szewczyk K, Lipinska B, Taylor A (1981) The R gene product of bacteriophage λ is the murein transglycosylase. Mol Gen Genet 184:111–114
Davis NG, Model P (1985) An artificial anchor domain: hydrophobicity suffices to stop transfer. Cell 41:607–614
Davis NG, Boeke JD, Model P (1985) Fine structure of a membrane anchor domain. J Mol Biol 181:111–121
Drexler K, Riede I, Henning U (1986) Morphogenesis of the long tail fibers of bacteriophage T2 involves proteolytic processing of the polypeptide (gene product 37) constituting the distal part of the fiber. J Mol Biol 191:267–272
Freudl R (1989) Insertion of peptides into cell-surface-exposed areas of the Escherichia coli OmpA protein does not interfere with export and membrane assembly. Gene 82:229–236
Freudl R, Schwarz H, Klose M, Movva RN, Henning U (1985) The nature of information, required for export and sorting, present within the outer membrane protein OmpA of Escherichia coli K-12. EMBO J 4:3593–3598
Garrett JM, Young R (1982) Lethal action of bacteriophage λ S gene. J Virol 44:886–892
Hawkes R, Viday E, Gordon J (1982) A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem 119:142–147
Henning U, Schwarz H, Chen R (1979) Radioimmunological screening method for specific membrane proteins. Anal Biochem 97:153–157
Jackson ME, Pratt JM (1987) An 18-amino acid amphiphilic helix forms the membrane-anchoring domain of the Escherichia coli penicillin-binding protein 5. Mol Microbiol 1:23–28
Jackson ME, Pratt JM (1988) Analysis of the membrane-binding domain of penicillin-binding protein 5 of Escherichia coli. Mol Microbiol 2:563–568
Josslin R (1970) The lysis mechanism of phage T4: mutants affecting lysis. Virology 40:719–726
Josslin R (1971) Physiological studies on the t gene defect in T4-infected Escherichia coli. Virology 44:101–107
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
MacIntyre S, Mutschler B, Henning U (1991) Requirement of the SecB chaperone for export of a non-secretory polypeptide in Escherichia coli. Mol Gen Genet 227:224–228
Manoil C (1991) Analysis of membrane protein topology using alkaline phosphatase and β-galactosidase gene fusions. Methods Cell Biol 34:61–75
Manoil C, Beckwith J (1985) TnphoA: A transposon probe for protein export signals. Proc Natl Acad Sci USA 82:8129–8133
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Montag D, Degen M, Henning U (1987a) Nucleotide sequence of gene t (lysis gene) of the E. coli phage T4. Nucleic Acids Res 15:6736
Montag D, Riede I, Eschbach M-L, Degen M, Henning U (1987b) Receptor-recognizing proteins of T-even type bacteriophages. Constant and hypervariable regions and an unusual case of evolution. J Mol Biol 196:165–174
Mukai G, Streisinger G, Miller N (1967) The mechanism of lysis in phage T4-infected cells. Virology 33:398–404
Osborn MJ, Gander JE, Parisi E, Carson J (1972) Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem 247:3962–3972
Phillips TA, Vaughn V, Bloch PL, Neidhardt FC (1987) Geneprotein index of Escherichia coli K-12, Edition 2. In: Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium, Cellular and molecular biology. American Society for Microbiology, Washington, DC, p 919–966
Raab R, Neal G, Garrett J, Grimaila R, Fusselman R, Young R (1986) Mutational analysis of bacteriophage lambda lysis gene S. J Bacteriol 167:1035–1042
Reader RW, Siminovitch L (1971) Lysis defective mutants of bacteriophage λ: on the role of the S function in lysis. Virology 143:280–289
Rennell D, Poteete AR (1985) Phage P22 lysis genes: nucleotide sequences and functional relationships with T4 and λ genes. Virology 143:280–289
Riede I (1987) Lysis gene t of T-even bacteriophages: Evidence that colicins and bacteriophage genes have common ancestors. J Bacteriol 169:2956–2961
Sakaguchi M, Mihara K, Sato R (1987) A short amino-terminal segment of microsomal cytochrome P-450 functions both as an insertion signal and as a stop-transfer sequence. EMBO J 6:2425–2431
Sanger F, Coulson AR, Hong GF, Hill DF, Petersen GB (1982) Nucleotide sequence of bacteriophage λ DNA. J Mol Biol 162:729–773
Schiffer M, Edmundson AB (1967) Use of helical wheels to represent the structures of proteins and to identify segments with helical potential. Biophys J 7:121–137
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Tsugita A, Inouye M, Terzaghi E, Streisinger G (1968) Purification of bacteriophage T4 lysozyme. J Biol Chem 234:391–397
von Heijne G (1985) Structural and thermodynamic aspects of the transfer of proteins into and across membranes. Curr Top Membr Transp 24:151–179
Wandersman C, Schwartz M, Ferenci T (1979) Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol 140:113
Wilson DB (1982) Effect of the λS gene product on properties of the Escherichia coli inner membrane. J Bacteriol 151:1403–1410
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors. Gene 33:103–119
Zagotta MT, Wilson DB (1990) Oligomerization of bacteriophage lambda S protein in the inner membrane of Escherichia coli. J Bacteriol 172:912–921
Author information
Authors and Affiliations
Additional information
Communicated by J. Lengeler
Rights and permissions
About this article
Cite this article
Lu, MJ., Henning, U. Lysis protein T of bacteriophage T4. Molec. Gen. Genet. 235, 253–258 (1992). https://doi.org/10.1007/BF00279368
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00279368