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. 1978 Aug;27(2):409–426. doi: 10.1128/jvi.27.2.409-426.1978

Isolation and characterization of bacteriophage T4 mutant preheads.

L Onorato, B Stirmer, M K Showe
PMCID: PMC354179  PMID: 691116

Abstract

To determine the function of individual gene products in the assembly and maturation of the T4 prehead, we have isolated and characterized aberrant preheads produced by mutations in three of the T4 head genes. Mutants in gene 21, which codes for the T4 maturation proteases, produce rather stable preheads whose morphology and protein composition are consistent with a wild-type prehead blocked in the maturation cleavages. Mutants in gene 24 produce similar structures which are unstable because they have gaps at all of their icosahedral vertices except the membrane attachment site. In addition, greatly elongated "giant preheads" are produced, suggesting that in the absence of P24 at the vertices, the distal cap of the prehead is unstable, allowing abnormal elongation of broth the prehead core and its shell. Vertex completion by P24 is required to allow the maturation cleavages to occur, and 24- preheads can be matured to capsids in vitro by the addition of P24. Preheads produced by a temperature-sensitive mutant in gene 23 are deficient in core proteins. We show that the shell of these preheads has the expanded lattice characteristic of the mature capsid as well as the binding sites for the proteins hoc and soc, even though none of the maturation cleavage takes place. We also show that 21- preheads composed of wild-type P23 can be expanded in vitro without cleavage.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aebi U., Bijlenga R., v d Broek J., v d Broek H., Eiserling F., Kellenberger C., Kellenberger E., Mesyanzhinov V., Müller L., Showe M. The transformation of tau particles into T4 heads. II. Transformations of the surface lattice and related observations on form determination. J Supramol Struct. 1974;2(2-4):253–275. doi: 10.1002/jss.400020218. [DOI] [PubMed] [Google Scholar]
  2. Bijlenga R. K., Aebi U., Kellenberger E. Properties and structure of a gene 24-controlled T4 giant phage. J Mol Biol. 1976 May 25;103(3):469–498. doi: 10.1016/0022-2836(76)90213-8. [DOI] [PubMed] [Google Scholar]
  3. Bijlenga R. K., Broek R vd, Kellenberger E. The transformation of rho-particles into T4 heads. I. Evidence for the conservative mode of this transformation. J Supramol Struct. 1974;2(1):45–59. doi: 10.1002/jss.400020106. [DOI] [PubMed] [Google Scholar]
  4. Favre R., Boy de la Tour E., Segrè N., Kellenberger E. Studies on the morphopoiesis of the head of phage T-even. I. Morphological, immunological, and genetic characterization of polyheads. J Ultrastruct Res. 1965 Oct;13(3):318–342. doi: 10.1016/s0022-5320(65)80080-6. [DOI] [PubMed] [Google Scholar]
  5. Fenner C., Traut R. R., Mason D. T., Wikman-Coffelt J. Quantification of Coomassie Blue stained proteins in polyacrylamide gels based on analyses of eluted dye. Anal Biochem. 1975 Feb;63(2):595–602. doi: 10.1016/0003-2697(75)90386-3. [DOI] [PubMed] [Google Scholar]
  6. Ishii T., Yanagida M. The two dispensable structural proteins (soc and hoc) of the T4 phage capsid; their purification and properties, isolation and characterization of the defective mutants, and their binding with the defective heads in vitro. J Mol Biol. 1977 Feb 5;109(4):487–514. doi: 10.1016/s0022-2836(77)80088-0. [DOI] [PubMed] [Google Scholar]
  7. Kellenberger E., Eiserling F. A., Boy de la Tour E. Studies on the morphopoiesis of the head of phage T-even. 3. The cores of head-related structures. J Ultrastruct Res. 1967 Dec 12;21(3):335–360. doi: 10.1016/s0022-5320(67)80099-6. [DOI] [PubMed] [Google Scholar]
  8. Kurtz M. B., Champe S. P. Precursors of the T4 internal peptides. J Virol. 1977 May;22(2):412–419. doi: 10.1128/jvi.22.2.412-419.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K., Eiserling F. A. Studies on the morphopoiesis of the head of phage T-even. V. The formation of polyheads. Mol Gen Genet. 1968 May 17;101(4):333–345. doi: 10.1007/BF00436231. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K., Johnson R. A. Maturation of the head of bacteriophage T4. II. Head-related, aberrant tau-particles. J Mol Biol. 1973 Nov 15;80(4):601–611. doi: 10.1016/0022-2836(73)90199-x. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K., Mölbert E., Showe M., Kellenberger E. Form-determining function of the genes required for the assembly of the head of bacteriophage T4. J Mol Biol. 1970 Apr 14;49(1):99–113. doi: 10.1016/0022-2836(70)90379-7. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K., Quittner S. F. Maturation of the head of bacteriophage T4. IV. The proteins of the core of the tubular polyheads and in vitro cleavage of the head proteins. Virology. 1974 Dec;62(2):483–499. doi: 10.1016/0042-6822(74)90409-7. [DOI] [PubMed] [Google Scholar]
  15. Luftig R. B., Lundh N. P. Bacteriophage T4 head morphogenesis. Isolation, partial characterization, and fate of gene 21-defective tau-particles. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1636–1640. doi: 10.1073/pnas.70.6.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McNicol L. A., Simon L. E. A mutation which bypasses the requirement for p24 in bacteriophage T4 capsid morphogenesis. J Mol Biol. 1977 Oct 25;116(2):261–283. doi: 10.1016/0022-2836(77)90216-9. [DOI] [PubMed] [Google Scholar]
  17. Müller-Salamin L., Onorato L., Showe M. K. Localization of minor protein components of the head of bacteriophage T4. J Virol. 1977 Oct;24(1):121–134. doi: 10.1128/jvi.24.1.121-134.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Onorato L., Showe M. K. Gene 21 protein-dependent proteolysis in vitro of purified gene 22 product of bacteriophage T4. J Mol Biol. 1975 Mar 5;92(3):395–412. doi: 10.1016/0022-2836(75)90288-0. [DOI] [PubMed] [Google Scholar]
  19. Paulson J. R., Laemmli U. K. Morphogenetic core of the bacteriophage T4 head. Structure of the core in polyheads. J Mol Biol. 1977 Apr 25;111(4):459–485. doi: 10.1016/s0022-2836(77)80064-8. [DOI] [PubMed] [Google Scholar]
  20. Showe M. K., Black L. W. Assembly core of bacteriophage T4: an intermediate in head formation. Nat New Biol. 1973 Mar 21;242(116):70–75. doi: 10.1038/newbio242070a0. [DOI] [PubMed] [Google Scholar]
  21. Showe M. K., Isobe E., Onorato L. Bacteriophage T4 prehead proteinase. I. Purification and properties of a bacteriophage enzyme which cleaves the capsid precursor proteins. J Mol Biol. 1976 Oct 15;107(1):35–54. doi: 10.1016/s0022-2836(76)80016-2. [DOI] [PubMed] [Google Scholar]
  22. Showe M. K., Isobe E., Onorato L. Bacteriophage T4 prehead proteinase. II. Its cleavage from the product of gene 21 and regulation in phage-infected cells. J Mol Biol. 1976 Oct 15;107(1):55–69. doi: 10.1016/s0022-2836(76)80017-4. [DOI] [PubMed] [Google Scholar]
  23. Simon L. D. Infection of Escherichia coli by T2 and T4 bacteriophages as seen in the electron microscope: T4 head morphogenesis. Proc Natl Acad Sci U S A. 1972 Apr;69(4):907–911. doi: 10.1073/pnas.69.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Steven A. C., Aebi U., Showe M. K. Folding and capsomere morphology of the P23 surface shell of bacteriophage T4 polyheads from mutants in five different head genes. J Mol Biol. 1976 Apr 15;102(3):373–400. [PubMed] [Google Scholar]
  25. van Driel R. Assembly of bacteriophage T4 head-related structures. Assembly of polyheads in vitro. J Mol Biol. 1977 Jul;114(1):61–72. doi: 10.1016/0022-2836(77)90283-2. [DOI] [PubMed] [Google Scholar]

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