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. 1976 Oct;128(1):130–143. doi: 10.1128/jb.128.1.130-143.1976

Independence of deoxyribonucleic acid replication and initiation from membrane fluidity and the supply of unsaturated fatty acids in Escherichia coli.

L Thilo, W Vielmetter
PMCID: PMC232835  PMID: 789329

Abstract

Mutant derivatives of the unsaturated fatty acid auxotroph K1062 were employed to investigate whether the supposedly membrane-bound bacterial replication machinery requires for its replicatory functions a fluid membrane environment as is known for several membrane-associated protein functions. Temperatures Tt for fluid reversible nonfluid phase transitions of membrane phospholipids are raised from below 18 to 38 degrees C when mutant cells are supplemented with elaidate instead of with oleate. In this experimental system current or synchroneously initiated new rounds of DNA replication are shown in vivo to continue 8 degrees below Tt, provided appropriate corrections for the concurrent cellular metabolic breakdown are considered. Temperature rate profiles for in vitro deoxyribonucleic acid replication rates measured in lysates of either oleate- or elaidate-supplemented cells yield congruent Arrhenius plots without discontinuities at corresponding Tt positions. We conclude that neither the start nor the propagation of replication forks depends on a fluid membrane. The capacity for the assembly of new replication complexes was studied in replication-aligned cells either shifted from oleate to elaidate (at temperatures below Tt for newly synthesized phospholipids) or starved for oleate. Regardless of whether unsaturated fatty acids are exchanged or completely withheld, new replication complexes can be normally assembled and initiated. These results do not support the conclusions reached by Fralick and Lark (1973) that the availability of unsaturated fatty acids is a prerequisite for the assembly of a functional replication complex.

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Selected References

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

  1. Anraku Y. The reduction and restoration of galactose transport in osmotically shocked cells of Escherichia coli. J Biol Chem. 1967 Mar 10;242(5):793–800. [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fielding P., Fox C. F. Evidence for stable attachment of DNA to membrane at the replication origin of Escherichia coli. Biochem Biophys Res Commun. 1970 Oct 9;41(1):157–162. doi: 10.1016/0006-291x(70)90482-1. [DOI] [PubMed] [Google Scholar]
  4. Fralick J. A., Lark K. G. Evidence for the involvement of unsaturated fatty acids in initiating chromosome replication in Escherichia coli. J Mol Biol. 1973 Nov 5;80(3):459–475. doi: 10.1016/0022-2836(73)90416-6. [DOI] [PubMed] [Google Scholar]
  5. Glaser M., Bayer W. H., Bell R. M., Vagelos P. R. Regulation of macromolecular biosynthesis in a mutant of Escherichia coli defective in membrane phospholipid biosynthesis. Proc Natl Acad Sci U S A. 1973 Feb;70(2):385–389. doi: 10.1073/pnas.70.2.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HANAWALT P. C., MAALOE O., CUMMINGS D. J., SCHAECHTER M. The normal DNA replication cycle. II. J Mol Biol. 1961 Apr;3:156–165. doi: 10.1016/s0022-2836(61)80042-9. [DOI] [PubMed] [Google Scholar]
  7. Hammer-Jespersen K., Munch-Petersen A., Schwartz M., Nygaard P. Induction of enzymes involed in the catabolism of deoxyribonucleosides and ribonucleosides in Escherichia coli K 12. Eur J Biochem. 1971 Apr 30;19(4):533–538. doi: 10.1111/j.1432-1033.1971.tb01345.x. [DOI] [PubMed] [Google Scholar]
  8. Henning U., Dennert G., Rehn K., Deppe G. Effects of oleate starvation in a fatty acid auxotroph of Escherichia coli K-12. J Bacteriol. 1969 May;98(2):784–796. doi: 10.1128/jb.98.2.784-796.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Herrmann R., Huf J., Bonhoeffer F. Cross hybridization and rate of chain elongation of the two classes of DNA intermediates. Nat New Biol. 1972 Dec 20;240(103):235–237. doi: 10.1038/newbio240235a0. [DOI] [PubMed] [Google Scholar]
  10. Jones N. C., Donachie W. D. Chromosome replication, transcription and control of cell division in Escherichia coli. Nat New Biol. 1973 May 23;243(125):100–103. [PubMed] [Google Scholar]
  11. Kass L. R., Bloch K. On the enzymatic synthesis of unsaturated fatty acids in Escherichia coli. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1168–1173. doi: 10.1073/pnas.58.3.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kass L. R. The antibacterial activity of 3-decynoyl-n-acetylcysteamine. Inhibition in vivo of beta-hydroxydecanoyl thioester dehydrase. J Biol Chem. 1968 Jun 25;243(12):3223–3228. [PubMed] [Google Scholar]
  13. Lark K. G. Regulation of chromosome replication and segregation in bacteria. Bacteriol Rev. 1966 Mar;30(1):3–32. doi: 10.1128/br.30.1.3-32.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Linden C. D., Wright K. L., McConnell H. M., Fox C. F. Lateral phase separations in membrane lipids and the mechanism of sugar transport in Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2271–2275. doi: 10.1073/pnas.70.8.2271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nunn W. D., Cronan J. E., Jr Unsaturated fatty acid synthesis is not required for induction of lactose transport in Escherichia coli. J Biol Chem. 1974 Feb 10;249(3):724–731. [PubMed] [Google Scholar]
  16. Overath P., Brenner M., Gulik-Krzywicki T., Shechter E., Letellier L. Lipid phase transitions in cytoplasmic and outer membranes of Escherichia coli. Biochim Biophys Acta. 1975 May 6;389(2):358–369. doi: 10.1016/0005-2736(75)90328-4. [DOI] [PubMed] [Google Scholar]
  17. Overath P., Hill F. F., Lamnek-Hirsch I. Biogenesis of E. coli membrane: evidence for randomization of lipid phase. Nat New Biol. 1971 Dec 29;234(52):264–267. doi: 10.1038/newbio234264a0. [DOI] [PubMed] [Google Scholar]
  18. Overath P., Schairer H. U., Stoffel W. Correlation of in vivo and in vitro phase transitions of membrane lipids in Escherichia coli. Proc Natl Acad Sci U S A. 1970 Oct;67(2):606–612. doi: 10.1073/pnas.67.2.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Overath P., Träuble H. Phase transitions in cells, membranes, and lipids of Escherichia coli. Detection by fluorescent probes, light scattering, and dilatometry. Biochemistry. 1973 Jul 3;12(14):2625–2634. doi: 10.1021/bi00738a012. [DOI] [PubMed] [Google Scholar]
  20. PRITCHARD R. H., LARK K. G. INDUCTION OF REPLICATION BY THYMINE STARVATION AT THE CHROMOSOME ORIGIN IN ESCHERICHIA COLI. J Mol Biol. 1964 Aug;9:288–307. doi: 10.1016/s0022-2836(64)80208-4. [DOI] [PubMed] [Google Scholar]
  21. Parker D. L., Glaser D. A. Chromosomal sites of DNA-membrane attachment in Escherichia coli. J Mol Biol. 1974 Aug 5;87(2):153–168. doi: 10.1016/0022-2836(74)90140-5. [DOI] [PubMed] [Google Scholar]
  22. Parker D. L., Glaser D. A. Effect of growth conditions on DNA-membrane attachment in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2446–2450. doi: 10.1073/pnas.72.6.2446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ryter A. Association of the nucleus and the membrane of bacteria: a morphological study. Bacteriol Rev. 1968 Mar;32(1):39–54. doi: 10.1128/br.32.1.39-54.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schaller H., Otto B., Nüsslein V., Huf J., Herrmann R., Bonhoeffer F. Deoxyribonucleic acid replication in vitro. J Mol Biol. 1972 Jan 28;63(2):183–200. doi: 10.1016/0022-2836(72)90369-5. [DOI] [PubMed] [Google Scholar]
  25. Schekman R., Weiner J. H., Weiner A., Kornberg A. Ten proteins required for conversion of phiX174 single-stranded DNA to duplex form in vitro. Resolution and reconstitution. J Biol Chem. 1975 Aug 10;250(15):5859–5865. [PubMed] [Google Scholar]
  26. Silbert D. F. Arrangement of fatty acyl groups in phosphatidylethanolamine from a fatty acid auxotroph of Escherichia coli. Biochemistry. 1970 Sep 1;9(18):3631–3640. doi: 10.1021/bi00820a021. [DOI] [PubMed] [Google Scholar]
  27. Silbert D. F., Ruch F., Vagelos P. R. Fatty acid replacements in a fatty acid auxotroph of Escherichia coli. J Bacteriol. 1968 May;95(5):1658–1665. doi: 10.1128/jb.95.5.1658-1665.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith D. W., Hanawalt P. C. Properties of the growing point region in the bacterial chromosome. Biochim Biophys Acta. 1967 Dec 19;149(2):519–531. doi: 10.1016/0005-2787(67)90180-3. [DOI] [PubMed] [Google Scholar]
  29. Sueoka N., Hammers J. M. Isolation of DNA-membrane complex in Bacillus subtilis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4787–4791. doi: 10.1073/pnas.71.12.4787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sueoka N., Quinn W. G. Membrane attachment of the chromosome replication origin in Bacillus subtilis. Cold Spring Harb Symp Quant Biol. 1968;33:695–705. doi: 10.1101/sqb.1968.033.01.078. [DOI] [PubMed] [Google Scholar]
  31. Thilo L., Overath P. Randomization of membrane lipids in relation to transport system assembly in Escherichia coli. Biochemistry. 1976 Jan 27;15(2):328–334. doi: 10.1021/bi00647a014. [DOI] [PubMed] [Google Scholar]
  32. Tsukagoshi N., Fox C. F. Abortive assembly of the lactose transport system in Escherichia coli. Biochemistry. 1973 Jul 17;12(15):2816–2822. doi: 10.1021/bi00739a007. [DOI] [PubMed] [Google Scholar]
  33. Wickner R. B., Hurwitz J. DNA replication in Escherichia coli made permeable by treatment with high sucrose. Biochem Biophys Res Commun. 1972 Apr 14;47(1):202–211. doi: 10.1016/s0006-291x(72)80029-9. [DOI] [PubMed] [Google Scholar]
  34. Wilson G., Fox C. F. Biogenesis of microbial transport systems: evidnce for coupled incorporation of newly synthesized lipids and proteins into membrane. J Mol Biol. 1971 Jan 14;55(1):49–60. doi: 10.1016/0022-2836(71)90280-4. [DOI] [PubMed] [Google Scholar]
  35. Worcel A., Burgi E. Properties of a membrane-attached form of the folded chromosome of Escherichia coli. J Mol Biol. 1974 Jan 5;82(1):91–105. doi: 10.1016/0022-2836(74)90576-2. [DOI] [PubMed] [Google Scholar]
  36. Yamaguchi K., Yoshikawa H. Association of the replication terminus of the Bacillus subtilis chromosome to the cell membrane. J Bacteriol. 1975 Nov;124(2):1030–1033. doi: 10.1128/jb.124.2.1030-1033.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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