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. 1974 Jun;77(2):199–220. doi: 10.1093/genetics/77.2.199

Phenethyl Alcohol Resistance in ESCHERICHIA COLI. III. a Temperature-Sensitive Mutation (dnaP) Affecting DNA Replication

Chieko Wada 1, Takashi Yura 1
PMCID: PMC1213124  PMID: 4603161

Abstract

A temperature-sensitive DNA replication mutant of E. coli K-12 was isolated among the mutants selected for phenethyl alcohol resistance at low temperatures. This mutation, designated as dnaP18, affects sensitivity of the cell to phenethyl alcohol, sodium deoxycholate and rifampicin, presumably due to an alteration in the membrane structure. At high temperatures (e.g., 42°), synthesis of DNA, but not RNA or protein, is arrested, leading to the formation of "filaments" in which no septum formation is apparent. Nucleoids observed under electron microscope seem to become dispersed and DNA fibrils less condensed, which may explain the loss of viability under these conditions. Genetic analyses, including reversion studies, indicate that a recessive dnaP mutation located between cya and metE on the chromosome is responsible for both alterations of the membrane properties and temperature sensitivity. The dnaP18 mutation does not affect growth of phage T4 or lambda under conditions where host DNA replication is completely inhibited. Kinetic studies of DNA replication and cell division in this mutant after the temperature shift from 30 to 42°, and during the subsequent recovery at 30°, accumulated evidence suggesting that DNA replication comes to a halt at 42° upon completion of a cycle already initiated before the temperature shift. Since the recovery of DNA synthesis after exposure to 42° does not depend on protein or RNA synthesis or other energy-requiring processes, the product of the mutant dnaP gene appears to be reversibly inactivated at 42°. Taken together with the recessive nature of the present mutation, it was suggested that one of the membrane proteins involved in initiation of DNA replication is affected in this mutant.

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Selected References

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  1. BONHOEFFER F., SCHALLER H. A METHOD FOR SELECTIVE ENRICHMENT OF MUTANTS BASED ON THE HIGH UV SENSITIVITY OF DNA CONTAINING 5-BROMOURACIL. Biochem Biophys Res Commun. 1965 Jun 18;20:93–97. [PubMed] [Google Scholar]
  2. Calendar R., Lindqvist B., Sironi G., Clark A. J. Characterization of REP- mutants and their interaction with P2 phage. Virology. 1970 Jan;40(1):72–83. doi: 10.1016/0042-6822(70)90380-6. [DOI] [PubMed] [Google Scholar]
  3. Carl P. L. Escherichia coli mutants with temperature-sensitive synthesis of DNA. Mol Gen Genet. 1970;109(2):107–122. doi: 10.1007/BF00269647. [DOI] [PubMed] [Google Scholar]
  4. Gross J. D. DNA replication in bacteria. Curr Top Microbiol Immunol. 1972;57:39–74. doi: 10.1007/978-3-642-65297-4_2. [DOI] [PubMed] [Google Scholar]
  5. Hirota Y., Mordoh J., Jacob F. On the process of cellular division in Escherichia coli. 3. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol. 1970 Nov 14;53(3):369–387. doi: 10.1016/0022-2836(70)90072-0. [DOI] [PubMed] [Google Scholar]
  6. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  7. Kohiyama M., Cousin D., Ryter A., Jacob F. Mutants thermosensibles d'Escherichia coli K 12. I. Isolement et caractérisation rapide. Ann Inst Pasteur (Paris) 1966 Apr;110(4):465–486. [PubMed] [Google Scholar]
  8. Kohiyama M. DNA synthesis in temperature sensitive mutants of Escherichia coli. Cold Spring Harb Symp Quant Biol. 1968;33:317–324. doi: 10.1101/sqb.1968.033.01.036. [DOI] [PubMed] [Google Scholar]
  9. Kuempel P. L. Temperature-sensitive initiation of chromosome replication in a mutant of Escherichia coli. J Bacteriol. 1969 Dec;100(3):1302–1310. doi: 10.1128/jb.100.3.1302-1310.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lark K. G., Lark C. Regulation of chromosome replication in Escherichia coli: a comparison of the effects of phenethyl alcohol treatment with those of amino acid starvation. J Mol Biol. 1966 Sep;20(1):9–19. doi: 10.1016/0022-2836(66)90113-6. [DOI] [PubMed] [Google Scholar]
  11. Nagel de Zwaig R., Luria S. E. Genetics and physiology of colicin-tolerant mutants of Escherichia coli. J Bacteriol. 1967 Oct;94(4):1112–1123. doi: 10.1128/jb.94.4.1112-1123.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nunn W. D., Tropp B. E. Effects of phenethyl alcohol on phospholipid metabolism in Escherichia coli. J Bacteriol. 1972 Jan;109(1):162–168. doi: 10.1128/jb.109.1.162-168.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ogawa H., Shimada K., Tomizawa J. Studies on radiation-sensitive mutants of E. coli. I. Mutants defective in the repair synthesis. Mol Gen Genet. 1968 May 3;101(3):227–244. doi: 10.1007/BF00271625. [DOI] [PubMed] [Google Scholar]
  14. RYTER A., KELLENBERGER E., BIRCHANDERSEN A., MAALOE O. Etude au microscope électronique de plasmas contenant de l'acide désoxyribonucliéique. I. Les nucléoides des bactéries en croissance active. Z Naturforsch B. 1958 Sep;13B(9):597–605. [PubMed] [Google Scholar]
  15. Schleif R. Isolation and characterization of streptolydigin resistant RNA polymerase. Nature. 1969 Sep 6;223(5210):1068–1069. doi: 10.1038/2231068a0. [DOI] [PubMed] [Google Scholar]
  16. Schubach W. H., Whitmer J. D., Davern C. I. Genetic control of DNA initiation in Escherichia coli. J Mol Biol. 1973 Feb 25;74(2):205–221. doi: 10.1016/0022-2836(73)90107-1. [DOI] [PubMed] [Google Scholar]
  17. Shapiro B. M., Siccardi A. G., Hirota Y., Jacob F. On the process of cellular division in Escherichia coli. Membrane prtein alterations associated with mutations affecting the initiation of DNA synthesis. J Mol Biol. 1970 Aug 28;52(1):75–89. doi: 10.1016/0022-2836(70)90178-6. [DOI] [PubMed] [Google Scholar]
  18. Siccardi A. G., Shapiro B. M. On the process of cellular division in Escherichia coli. IV. Altered protein composition and turnover of the membranes of thermosensitive mutants defective in chromosomal replication. J Mol Biol. 1971 Mar 28;56(3):475–490. doi: 10.1016/0022-2836(71)90395-0. [DOI] [PubMed] [Google Scholar]
  19. Wada J. A. Correlative behavioral and evoked-potential analysis of "inhibitory" temporal-lobe function. Recent Adv Biol Psychiatry. 1968;10:220–239. doi: 10.1007/978-1-4684-9072-5_17. [DOI] [PubMed] [Google Scholar]
  20. Yura T., Wada C. Phenethyl alcohol resistance in Escherichia coli. I. Resistance of strain C600 and its relation to azide resistance. Genetics. 1968 Jun;59(2):177–190. doi: 10.1093/genetics/59.2.177. [DOI] [PMC free article] [PubMed] [Google Scholar]

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