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. 1977 Oct;24(1):363–377. doi: 10.1128/jvi.24.1.363-377.1977

Morphogenesis of Bacteriophage φ29 of Bacillus subtilis: Mapping and Functional Analysis of the Head Fiber Gene

Bernard E Reilly 1, Rodger A Nelson 1, Dwight L Anderson 1
PMCID: PMC515937  PMID: 409854

Abstract

A set of mutants of Bacillus subtilis bacteriophage φ29 unable to synthesize the head fiber protein has been identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Infectious phage are produced during restrictive infection. We have focused on mutant sus 8.5(900) because the mutation is suppressible by both the su+3 and su+44 hosts, and it can be mapped by three- and four-factor crosses. After restrictive infection with mutant sus 8.5(900), a fragment about 70% of the size of the normal fiber is produced as well as particles that are fast-sedimenting in sucrose gradients relative to φ29+. These particles have the buoyant density of particles with the fibers removed and have the absolute plating efficiency of φ29+. Fiber protein is absent from prohead as well as virion. A second set of mutants produces fiber protein with a slightly altered electrophoretic mobility. This type of fiber protein is either present or absent on both prohead and virion. A third class of mutants, typified by 914, produces a “normal” fiber, but a major head protein of altered electrophoretic mobility. After infection by this mutant, the fiber is absent from both prohead and virion, and the biological and physical properties of the 914− particle are similar to those of particles produced after infection of the su− host by sus8.5(900). These observations suggest that the head fiber is not an essential component of the prohead or virion and that the assembly process is efficient in the absence of fiber protein. Three- and four-factor genetic crosses have established the order sus8(769)-8(914)-sus8.5(900)-sus9(756) and indicate that cistrons 8 and 8.5 code for the major head protein and head fiber protein, respectively.

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Selected References

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

  1. Anderson D. L., Hickman D. D., Reilly B. E. Structure of Bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid. J Bacteriol. 1966 May;91(5):2081–2089. doi: 10.1128/jb.91.5.2081-2089.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson D. L., Mosharrafa E. T. Physical and biological properties of phage phi 29 deoxyribonucleic acid. J Virol. 1968 Oct;2(10):1185–1190. doi: 10.1128/jvi.2.10.1185-1190.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson D. L., Pollock M. E., Brower L. F. Morphology of Mycoplasma laidlawii type A. I. Comparison of electron microscopic counts with colony-forming units. J Bacteriol. 1965 Dec;90(6):1764–1767. doi: 10.1128/jb.90.6.1764-1767.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson D. L., Reilly B. E. Analysis of bacteriophage phi 29 gene function: protein synthesis in suppressor-sensitive mutant infection of Bacillus subtilis. J Virol. 1974 Jan;13(1):211–221. doi: 10.1128/jvi.13.1.211-221.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Black L. W. Bacteriophage T4 internal protein mutants: isolation and properties. Virology. 1974 Jul;60(1):166–179. doi: 10.1016/0042-6822(74)90374-2. [DOI] [PubMed] [Google Scholar]
  6. Camacho A., Moreno F., Carrascosa J. L., Viñuela E., Salas M. A suppressor of nonsense mutations in Bacillus subtilis. Eur J Biochem. 1974 Aug 15;47(1):199–205. doi: 10.1111/j.1432-1033.1974.tb03683.x. [DOI] [PubMed] [Google Scholar]
  7. Carrascosa J. L., Camacho A., Moreno F., Jiménez F., Mellado R. P., Viñuela E., Salas M. Bacillus subtilis phage phi29. Characterization of gene products and functions. Eur J Biochem. 1976 Jul 1;66(2):229–241. doi: 10.1111/j.1432-1033.1976.tb10512.x. [DOI] [PubMed] [Google Scholar]
  8. Carrascosa J. L., Viñuela E., Salas M. Proteins induced in Bacillus subtilis infected with bacteriophage phi 29. Virology. 1973 Nov;56(1):291–299. [PubMed] [Google Scholar]
  9. Hagen E. W., Reilly B. E., Tosi M. E., Anderson D. L. Analysis of gene function of bacteriophage phi 29 of Bacillus subtilis: identification of cistrons essential for viral assembly. J Virol. 1976 Aug;19(2):501–517. doi: 10.1128/jvi.19.2.501-517.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hawley L. A., Reilly B. E., Hagen E. W., Anderson D. L. Viral protein synthesis in bacteriophage phi 29-infected Bacillus subtilis. J Virol. 1973 Nov;12(5):1149–1159. doi: 10.1128/jvi.12.5.1149-1159.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ishii T., Yanagida M. Molecular organization of the shell of the Teven bacteriophage head. J Mol Biol. 1975 Oct 5;97(4):655–660. doi: 10.1016/s0022-2836(75)80065-9. [DOI] [PubMed] [Google Scholar]
  12. Jiménez F., Camacho A., De La Torre J., Viñuela E., Salas M. Assembly of Bacillus subtilis phage phe29. 2. Mutants in the cistrons coding for the non-structural proteins. Eur J Biochem. 1977 Feb 15;73(1):57–72. doi: 10.1111/j.1432-1033.1977.tb11291.x. [DOI] [PubMed] [Google Scholar]
  13. McGuire J. C., Pène J. J., Barrow-Carraway J. Gene expression during the development of bacteriophage phi 29. 3. Analysis of viral-specific protein synthesis with suppressible mutants. J Virol. 1974 Mar;13(3):690–698. doi: 10.1128/jvi.13.3.690-698.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mellado R. P., Moreno F., Viñuela E., Salas M., Reilly B. E., Anderson D. L. Genetic analysis of bacteriophage phi 29 of Bacillus subtilis: integration and mapping of reference mutants of two collections. J Virol. 1976 Aug;19(2):495–500. doi: 10.1128/jvi.19.2.495-500.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mellado R. P., Méndez E., Viñuela E., Salas M. Order of the two major head protein genes of bacteriophage phi 29 of Bacillus subtilis. J Virol. 1977 Oct;24(1):378–382. doi: 10.1128/jvi.24.1.378-382.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mellado R. P., Vinuela E., Salas M. Isolation of a strong suppressor of nonsense mutations in Bacillus subtilis. Eur J Biochem. 1976 May 17;65(1):213–223. doi: 10.1111/j.1432-1033.1976.tb10408.x. [DOI] [PubMed] [Google Scholar]
  17. Moreno F. Suppressor-sensitive mutants and genetic map of Bacillus subtilis bacteriophage phi 29. Virology. 1974 Nov;62(1):1–16. doi: 10.1016/0042-6822(74)90298-0. [DOI] [PubMed] [Google Scholar]
  18. Méndez E., Ramírez G., Salas M., Viñuela E. Structural proteins of bacteriophage phi 29. Virology. 1971 Sep;45(3):567–576. doi: 10.1016/0042-6822(71)90172-3. [DOI] [PubMed] [Google Scholar]
  19. Nelson R. A., Reilly B. E., Anderson D. L. Morphogenesis of bacteriophage phi 29 of Bacillus subtilis: preliminary isolation and characterization of intermediate particles of the assembly pathway. J Virol. 1976 Aug;19(2):518–532. doi: 10.1128/jvi.19.2.518-532.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ortin J., Viñuela E., Salas M., Vasquez C. DNA-protein complex in circular DNA from phage phi-29. Nat New Biol. 1971 Dec 29;234(52):275–277. doi: 10.1038/newbio234275a0. [DOI] [PubMed] [Google Scholar]
  21. Polsinelli M., Beretta M. Genetic Recombination in Crosses Between Streptomyces aureofaciens and Streptomyces rimosus. J Bacteriol. 1966 Jan;91(1):63–68. doi: 10.1128/jb.91.1.63-68.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Péne J. J., Murr P. C., Barrow-Carraway J. Synthesis of bacteriophage phi 29 proteins in Bacillus subtilis. J Virol. 1973 Jul;12(1):61–67. doi: 10.1128/jvi.12.1.61-67.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. REILLY B. E., SPIZIZEN J. BACTERIOPHAGE DEOXYRIBONUCLEATE INFECTION OF COMPETENT BACILLUS SUBTILIS. J Bacteriol. 1965 Mar;89:782–790. doi: 10.1128/jb.89.3.782-790.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reilly B. E., Tosi M. E., Anderson D. L. Genetic analysis of bacteriophage phi29 of Bacillus subtilis: mapping of the cistrons coding for structural proteins. J Virol. 1975 Oct;16(4):1010–1016. doi: 10.1128/jvi.16.4.1010-1016.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reilly B. E., Zeece V. M., Anderson D. L. Genetic study of suppressor-sensitive mutants of the Bacillus subtilis bacteriophage phi 29. J Virol. 1973 May;11(5):756–760. doi: 10.1128/jvi.11.5.756-760.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Salas M., Vásquez C., Méndez E., Viñuela E. Head fibers of bacteriophage phi 29. Virology. 1972 Oct;50(1):180–188. doi: 10.1016/0042-6822(72)90358-3. [DOI] [PubMed] [Google Scholar]
  27. Tosi M. E., Reilly B. E., Anderson D. L. Morphogenesis of bacteriophage phi29 of Bacillus subtilis: cleavage and assembly of the neck appendage protein. J Virol. 1975 Nov;16(5):1282–1295. doi: 10.1128/jvi.16.5.1282-1295.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tosi M., Anderson D. L. Antigenic properties of bacteriophage phi 29 structural proteins. J Virol. 1973 Dec;12(6):1548–1559. doi: 10.1128/jvi.12.6.1548-1559.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Viñuela E., Camacho A., Jiménez F., Carrascosa J. L., Ramírez G., Salas M. Structure and assembly of phage phi29. Philos Trans R Soc Lond B Biol Sci. 1976 Nov 30;276(943):29–35. doi: 10.1098/rstb.1976.0095. [DOI] [PubMed] [Google Scholar]
  30. Yanofsky S., Kawamura F., Ito J. Thermolabile transfecting DNA from temperature-sensitive mutant of phage phi29. Nature. 1976 Jan 1;259(5538):60–63. doi: 10.1038/259060a0. [DOI] [PubMed] [Google Scholar]

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