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. 1974 Nov;14(5):1115–1125. doi: 10.1128/jvi.14.5.1115-1125.1974

Cold-Sensitive Mutants of Bacteriophage φX174. II. Comparison of Two Cold-Sensitive Mutants

David J Segal a,1, Clifton E Dowell b
PMCID: PMC355628  PMID: 4610177

Abstract

Cold-sensitive bacteriophage φX174 mutants, another class of conditional lethals, were examined with regard to growth parameters, DNA synthesis, and particle properties. Two mutants, cs70 and cs82, were examined. Mutant cs70 was eclipse defective, showing altered eclipse kinetics at permissive temperature (40 C) and failing entirely to eclipse at restrictive temperature (25 C). Mutant cs70 replicated well at 25 C if allowed prior eclipse at 40 C. Mutant cs82 had wild-type eclipse at both temperatures but was defective in single-strand synthesis at 25 C, which led to delayed progeny phage appearance, decreased progeny phage synthesis rate, and greatly reduced burst size. The cs82 block could not be bypassed by temperature shift. Since complementation analysis of cs70 and cs82 was not feasible due to the unique properties of these mutants, those φX174 properties affected by the virus coat were examined as an index of a mutation in a coat protein gene. Mutant cs70 had aberrant attachment kinetics at both 25 C and 40 C, evidence of a coat protein alteration. Mutant cs70 also exhibited significantly decreased thermal stability, further evidence of an altered virus structure. Mutant cs82 had increased thermal stability, but the difference was not sufficient to allow unequivocal assignment of this mutant to a coat protein gene. Both mutants had wild-type antiserum inactivation and host range, although cs70 was subject to less of (low-level) plating restriction by endogenous F+ factors.

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Selected References

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

  1. Baker R., Tessman I. Heat stability of mutants in genes II, 3a, and VI of phage S13. Virology. 1968 May;35(1):179–181. doi: 10.1016/0042-6822(68)90321-8. [DOI] [PubMed] [Google Scholar]
  2. Bayer M. E., Starkey T. W. The adsorption of bacteriophage phi X174 and its interaction with Escherichia coli; a kinetic and morphological study. Virology. 1972 Jul;49(1):236–256. doi: 10.1016/s0042-6822(72)80026-6. [DOI] [PubMed] [Google Scholar]
  3. Benbow R. M., Hutchison C. A., Fabricant J. D., Sinsheimer R. L. Genetic Map of Bacteriophage phiX174. J Virol. 1971 May;7(5):549–558. doi: 10.1128/jvi.7.5.549-558.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benbow R. M., Mayol R. F., Picchi J. C., Sinsheimer R. L. Direction of Translation and Size of Bacteriophage phiX174 Cistrons. J Virol. 1972 Jul;10(1):99–114. doi: 10.1128/jvi.10.1.99-114.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bleichrodt J. F., Blok J., Berends-Van Abkoude E. R. Thermal inactivation of bacteriophage phi X174 and two of its mutants. Virology. 1968 Nov;36(3):343–355. doi: 10.1016/0042-6822(68)90160-8. [DOI] [PubMed] [Google Scholar]
  6. Bone D. R., Dowel C. E. A mutant of bacteriophage øX174 which infects E. coli K12 strains. I. Isolation and partial characterization of øXtB. Virology. 1973 Apr;52(2):319–329. [PubMed] [Google Scholar]
  7. Borrias W. E., van de Pol J. H., van de Vate C., van Arkel G. A. Complementation experiments between conditional lethal mutants of bacteriophage psiX 174. Mol Gen Genet. 1969 Oct 13;105(2):152–163. doi: 10.1007/BF00445685. [DOI] [PubMed] [Google Scholar]
  8. Bowes J. M., Dowell C. E. Purification and some properties of bacteriophage ST-1. J Virol. 1974 Jan;13(1):53–61. doi: 10.1128/jvi.13.1.53-61.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brown L. R., Dowell C. E. Replication of coliphage M-13. II. Intracellular deoxyribonucleic acid forms associated with M-13 infection of mitomycin C-treated cells. J Virol. 1968 Nov;2(11):1296–1307. doi: 10.1128/jvi.2.11.1296-1307.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. CAMPBELL A. Sensitive mutants of bacteriophage lambda. Virology. 1961 May;14:22–32. doi: 10.1016/0042-6822(61)90128-3. [DOI] [PubMed] [Google Scholar]
  11. Denhardt D. T. Formation of ribosylthymine in Escherichia coli. Studies on pulse labeling with thymine and thymidine. J Biol Chem. 1969 May 25;244(10):2710–2715. [PubMed] [Google Scholar]
  12. Denhardt D. T., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. 3. Phage maturation and lysis after synchronized infection. J Mol Biol. 1965 Jul;12(3):641–646. doi: 10.1016/s0022-2836(65)80318-7. [DOI] [PubMed] [Google Scholar]
  13. Doniger J., Tessman I. An S13 capsid mutant that makes no replicative-form DNA. Virology. 1969 Nov;39(3):389–394. doi: 10.1016/0042-6822(69)90086-5. [DOI] [PubMed] [Google Scholar]
  14. Dowell C. E. Cold-sensitive mutants of bacteriophage phi-X-174. I. A mutant blocked in the eclipse function at low temperature. Proc Natl Acad Sci U S A. 1967 Sep;58(3):958–961. doi: 10.1073/pnas.58.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dowell C. E., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. IX. Studies on the physiology of three phi-X174 temperature-sensitive mutants. J Mol Biol. 1966 Apr;16(2):374–386. doi: 10.1016/s0022-2836(66)80180-8. [DOI] [PubMed] [Google Scholar]
  16. FIERS W., SINSHEIMER R. L. The structure of the DNA of bacteriophage phi-X174. II. Thermal inactivation. J Mol Biol. 1962 Oct;5:420–423. doi: 10.1016/s0022-2836(62)80030-8. [DOI] [PubMed] [Google Scholar]
  17. FUJIMURA R., KAESBERG P. The adsorption of bacteriophage phi-X174 to its host. Biophys J. 1962 Nov;2:433–449. doi: 10.1016/s0006-3495(62)86866-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Forsheit A. B., Ray D. S. Conformations of the single-stranded DNA of bacteriophage M13. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1534–1541. doi: 10.1073/pnas.67.3.1534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Funk F. D., Sinsheimer R. L. Process of infection with bacteriophage phiX174. XXXV. Cistron 8. J Virol. 1970 Jul;6(1):12–19. doi: 10.1128/jvi.6.1.12-19.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. GUTHRIE G. D., SINSHEIMER R. L. Observations on the infection of bacterial protoplasts with the deoxyribonucleic acid of bacteriophage phi X174. Biochim Biophys Acta. 1963 Jun 25;72:290–297. [PubMed] [Google Scholar]
  21. Greenlee L. L., Sinsheimer R. L. The process of infection with bacteriophage phi X174. XVII. Effects of specific metabolic interruptions. J Mol Biol. 1968 Mar 14;32(2):303–320. doi: 10.1016/0022-2836(68)90011-9. [DOI] [PubMed] [Google Scholar]
  22. Groman N. B. Restriction of bacteriophage phiX174 by F factor. Biochem Biophys Res Commun. 1969 Nov 6;37(4):691–696. doi: 10.1016/0006-291x(69)90866-3. [DOI] [PubMed] [Google Scholar]
  23. Hutchison C. A., 3rd, Edgell M. H., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. XII. Phenotypic mixing between electrophoretic mutants of phi-X174. J Mol Biol. 1967 Feb 14;23(3):553–575. doi: 10.1016/s0022-2836(67)80125-6. [DOI] [PubMed] [Google Scholar]
  24. Iwaya M., Denhardt D. T. The mechanism of replication of phi X174 single-stranded DNA. II. The role of viral proteins. J Mol Biol. 1971 Apr 28;57(2):159–175. doi: 10.1016/0022-2836(71)90339-1. [DOI] [PubMed] [Google Scholar]
  25. Lindqvist B. H., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. XV. Bacteriophage DNA synthesis in abortive infections with a set of conditional lethal mutants. J Mol Biol. 1967 Nov 28;30(1):69–80. doi: 10.1016/0022-2836(67)90244-6. [DOI] [PubMed] [Google Scholar]
  26. Newbold J. E., Sinsheimer R. L. Process of infection with bacteriophage phi-X174. XXXIV. Kinetic of the attachment and eclipse steps of the infection. J Virol. 1970 Apr;5(4):427–431. doi: 10.1128/jvi.5.4.427-431.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. PFEIFER D. [Genetic studies on the bacteriophage phi-X174. I. Development of a selective system and demonstration of a genetic recombination]. Z Vererbungsl. 1961;92:317–329. [PubMed] [Google Scholar]
  28. Primrose S. B., Landers W. M., Dowell C. E. Replication of bacteriophage phiX174 in a temperature-sensitive deoxyribonucleic acid host cell. J Virol. 1971 Oct;8(4):594–596. doi: 10.1128/jvi.8.4.594-596.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. ROLFE U., SINSHEIMER R. L. ANTIGENS OF BACTERIOPHAGE PHI-X174. J Immunol. 1965 Jan;94:18–21. [PubMed] [Google Scholar]
  30. Siegel J. E., Hayashi M. N., Hayashi M. Phi-X-174 coat protein mutants affecting DNA synthesis. Biochem Biophys Res Commun. 1968 Jun 10;31(5):774–778. doi: 10.1016/0006-291x(68)90629-3. [DOI] [PubMed] [Google Scholar]
  31. Siegel J. E., Hayashi M. Phi-X-174 bacteriophage structural mutants which affect deoxyribonucleic acid synthesis. J Virol. 1969 Oct;4(4):400–407. doi: 10.1128/jvi.4.4.400-407.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sinsheimer R. L. Bacteriophage phi-X174 and related viruses. Prog Nucleic Acid Res Mol Biol. 1968;8:115–169. [PubMed] [Google Scholar]
  33. TESSMAN E. S. COMPLEMENTATION GROUPS IN PHAGE S13. Virology. 1965 Feb;25:303–321. doi: 10.1016/0042-6822(65)90208-4. [DOI] [PubMed] [Google Scholar]
  34. Tessman I., Tessman E. S. Functional units of phage S13: identification of two genes that determine the structure of the phage coat. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1459–1462. doi: 10.1073/pnas.55.6.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weisbeek P. J., van de Pol J. H., van Arkel G. A. Mapping of host range mutants of bacteriophage phiX174. Virology. 1973 Apr;52(2):408–416. doi: 10.1016/0042-6822(73)90335-8. [DOI] [PubMed] [Google Scholar]
  36. Zuccarelli A. J., Benbow R. M., Sinsheimer R. L. Deletion mutants of bacteriophage phiX174. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1905–1910. doi: 10.1073/pnas.69.7.1905. [DOI] [PMC free article] [PubMed] [Google Scholar]

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