Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1989 Jan;8(1):317–323. doi: 10.1002/j.1460-2075.1989.tb03379.x

The Escherichia coli lov gene product connects peptidoglycan synthesis, ribosomes and growth rate.

P Bouloc 1, A Jaffé 1, R D'Ari 1
PMCID: PMC400806  PMID: 2540960

Abstract

Mecillinam, a beta-lactam antibiotic which binds specifically to penicillin-binding protein 2 (PBP2), blocks lateral cell-wall elongation, induces spherical morphology and ultimately kills bacteria. We describe here a new mecillinam-resistant mutant of Escherichia coli, the lov mutant. It possesses active PBP2, as evidenced by its rod shape in the absence of mecillinam (but not in its presence), its ability to filament when septation is inhibited, and its penicillin-binding ability. The lov mutant grows slowly but seems to regulate its macromolecular parameters properly: cell volume, RNA content (ribosome concentration), and DNA content are appropriate for the growth rate, and the growth yield is identical to that of wild type. The lov mutation is located at 41 min on the E.coli genetic map and is recessive. Certain rpsL (StrR) mutations suppress the lov mutant's mecillinam resistance. The allele specificity of the suppression suggests that the lov gene product may interact directly with the ribosomes. The lov gene product thus seems to define a link between PBP2 (the mecillinam target) and the ribosomes; we propose that this link is involved in transmitting information on the growth rate (ribosome concentration) to the peptidoglycan synthesizing apparatus.

Full text

PDF
317

Selected References

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

  1. Antón D. N., de Micheli A. T., Palermo A. M. Isolation of round-cell mutants of Salmonella typhimurium. Can J Microbiol. 1983 Feb;29(2):170–173. doi: 10.1139/m83-029. [DOI] [PubMed] [Google Scholar]
  2. Aono R., Yamasaki M., Tamura G. High and selective resistance to mecillinam in adenylate cyclase-deficient or cyclic adenosine 3',5'-monophosphate receptor protein-deficient mutants of Escherichia coli. J Bacteriol. 1979 Feb;137(2):839–845. doi: 10.1128/jb.137.2.839-845.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Begg K. J., Donachie W. D. Cell shape and division in Escherichia coli: experiments with shape and division mutants. J Bacteriol. 1985 Aug;163(2):615–622. doi: 10.1128/jb.163.2.615-622.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bohman K., Ruusala T., Jelenc P. C., Kurland C. G. Kinetic impairment of restrictive streptomycin-resistant ribosomes. Mol Gen Genet. 1984;198(2):90–99. doi: 10.1007/BF00328706. [DOI] [PubMed] [Google Scholar]
  6. Bouloc P., Jaffé A., D'Ari R. Preliminary physiologic characterization and genetic analysis of a new Escherichia coli mutant, lov, resistant to mecillinam. Rev Infect Dis. 1988 Jul-Aug;10(4):905–910. doi: 10.1093/clinids/10.4.905. [DOI] [PubMed] [Google Scholar]
  7. Breckenridge L., Gorini L. Genetic analysis of streptomycin resistance in Escherichia coli. Genetics. 1970 May;65(1):9–25. doi: 10.1093/genetics/65.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cooper S., Helmstetter C. E. Chromosome replication and the division cycle of Escherichia coli B/r. J Mol Biol. 1968 Feb 14;31(3):519–540. doi: 10.1016/0022-2836(68)90425-7. [DOI] [PubMed] [Google Scholar]
  9. D'Ari R., Jaffé A., Bouloc P., Robin A. Cyclic AMP and cell division in Escherichia coli. J Bacteriol. 1988 Jan;170(1):65–70. doi: 10.1128/jb.170.1.65-70.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dombou M., Mizuno T., Mizushima S. Interaction of the cytoplasmic membrane and ribosomes in Escherichia coli; altered ribosomal proteins in sucrose-dependent spectinomycin-resistant mutants. Mol Gen Genet. 1977 Sep 21;155(1):53–60. doi: 10.1007/BF00268560. [DOI] [PubMed] [Google Scholar]
  11. Fraenkel D. G., Banerjee S. A mutation increasing the amount of a constitutive enzyme in Escherichia coli, glucose 6-phosphate dehydrogenase. J Mol Biol. 1971 Feb 28;56(1):183–194. doi: 10.1016/0022-2836(71)90093-3. [DOI] [PubMed] [Google Scholar]
  12. Frandsen E. K., Krishna G. A simple ultrasensitive method for the assay of cyclic AMP and cyclic GMP in tissues. Life Sci. 1976 Mar 1;18(5):529–541. doi: 10.1016/0024-3205(76)90331-3. [DOI] [PubMed] [Google Scholar]
  13. Guiso N., Blazy B. Regulatory aspects of the cyclic AMP receptor protein in Escherichia coli K-12. Biochem Biophys Res Commun. 1980 May 14;94(1):278–283. doi: 10.1016/s0006-291x(80)80217-8. [DOI] [PubMed] [Google Scholar]
  14. Ishino F., Matsuhashi M. Peptidoglycan synthetic enzyme activities of highly purified penicillin-binding protein 3 in Escherichia coli: a septum-forming reaction sequence. Biochem Biophys Res Commun. 1981 Aug 14;101(3):905–911. doi: 10.1016/0006-291x(81)91835-0. [DOI] [PubMed] [Google Scholar]
  15. Ishino F., Park W., Tomioka S., Tamaki S., Takase I., Kunugita K., Matsuzawa H., Asoh S., Ohta T., Spratt B. G. Peptidoglycan synthetic activities in membranes of Escherichia coli caused by overproduction of penicillin-binding protein 2 and rodA protein. J Biol Chem. 1986 May 25;261(15):7024–7031. [PubMed] [Google Scholar]
  16. Iwaya M., Jones C. W., Khorana J., Strominger J. L. Mapping of the mecillinam-resistant, round morphological mutants of Escherichia coli. J Bacteriol. 1978 Jan;133(1):196–202. doi: 10.1128/jb.133.1.196-202.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jaffé A., Chabbert Y. A., Derlot E. Selection and characterization of beta-lactam-resistant Escherichia coli K-12 mutants. Antimicrob Agents Chemother. 1983 Apr;23(4):622–625. doi: 10.1128/aac.23.4.622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jaffé A., D'Ari R., Norris V. SOS-independent coupling between DNA replication and cell division in Escherichia coli. J Bacteriol. 1986 Jan;165(1):66–71. doi: 10.1128/jb.165.1.66-71.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jaffé A., D'Ari R. Regulation of chromosome segregation in Escherichia coli. Ann Inst Pasteur Microbiol. 1985 Jan-Feb;136A(1):159–164. doi: 10.1016/s0769-2609(85)80036-3. [DOI] [PubMed] [Google Scholar]
  20. James R., Haga J. Y., Pardee A. B. Inhibition of an early event in the cell division cycle of Escherichia coli by FL1060, an amidinopenicillanic acid. J Bacteriol. 1975 Jun;122(3):1283–1292. doi: 10.1128/jb.122.3.1283-1292.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Joseleau-Petit D., Kepes F., Kepes A. Cyclic changes of the rate of phospholipid synthesis during synchronous growth of Escherichia coli. Eur J Biochem. 1984 Mar 15;139(3):605–611. doi: 10.1111/j.1432-1033.1984.tb08047.x. [DOI] [PubMed] [Google Scholar]
  22. Joseleau-Petit D., Képès F., Peutat L., D'Ari R., Képès A. DNA replication initiation, doubling of rate of phospholipid synthesis, and cell division in Escherichia coli. J Bacteriol. 1987 Aug;169(8):3701–3706. doi: 10.1128/jb.169.8.3701-3706.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  24. Lindahl L., Zengel J. M. Ribosomal genes in Escherichia coli. Annu Rev Genet. 1986;20:297–326. doi: 10.1146/annurev.ge.20.120186.001501. [DOI] [PubMed] [Google Scholar]
  25. Long W. S., Slayman C. L., Low K. B. Production of giant cells of Escherichia coli. J Bacteriol. 1978 Feb;133(2):995–1007. doi: 10.1128/jb.133.2.995-1007.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lund F., Tybring L. 6 -amidinopenicillanic acids--a new group of antibiotics. Nat New Biol. 1972 Apr 5;236(66):135–137. doi: 10.1038/newbio236135a0. [DOI] [PubMed] [Google Scholar]
  27. Matsuhashi S., Kamiryo T., Blumberg P. M., Linnett P., Willoughby E., Strominger J. L. Mechanism of action and development of resistance to a new amidino penicillin. J Bacteriol. 1974 Feb;117(2):578–587. doi: 10.1128/jb.117.2.578-587.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miyoshi Y., Yamagata H. Sucrose-dependent spectinomycin-resistant mutants of Escherichia coli. J Bacteriol. 1976 Jan;125(1):142–148. doi: 10.1128/jb.125.1.142-148.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mizuno T., Yamada H., Yamagata H., Mizushima S. Coordinated alterations in ribosomes and cytoplasmic membrane in sucrose-dependent, spectinomycin-resistant mutants of Escherichia coli. J Bacteriol. 1976 Feb;125(2):524–530. doi: 10.1128/jb.125.2.524-530.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mizuno T., Yamagata H., Mizushima S. Interaction of cytoplasmic membrane and ribosomes in Escherichia coli: spectinomycin-induced disappearance of membrane protein I-19. J Bacteriol. 1977 Jan;129(1):326–332. doi: 10.1128/jb.129.1.326-332.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Novel M., Novel G. Regulation of beta-glucuronidase synthesis in Escherichia coli K-12: constitutive mutants specifically derepressed for uidA expression. J Bacteriol. 1976 Jul;127(1):406–417. doi: 10.1128/jb.127.1.406-417.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Park J. T., Burman L. FL-1060: a new penicillin with a unique mode of action. Biochem Biophys Res Commun. 1973 Apr 16;51(4):863–868. doi: 10.1016/0006-291x(73)90006-5. [DOI] [PubMed] [Google Scholar]
  33. Pierucci O. Phospholipid synthesis during the cell division cycle of Escherichia coli. J Bacteriol. 1979 May;138(2):453–460. doi: 10.1128/jb.138.2.453-460.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Remaut E., Tsao H., Fiers W. Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication. Gene. 1983 Apr;22(1):103–113. doi: 10.1016/0378-1119(83)90069-0. [DOI] [PubMed] [Google Scholar]
  35. Shurvinton C. E., Lloyd R. G., Benson F. E., Attfield P. V. Genetic analysis and molecular cloning of the Escherichia coli ruv gene. Mol Gen Genet. 1984;194(1-2):322–329. doi: 10.1007/BF00383535. [DOI] [PubMed] [Google Scholar]
  36. Spratt B. G., Boyd A., Stoker N. Defective and plaque-forming lambda transducing bacteriophage carrying penicillin-binding protein-cell shape genes: genetic and physical mapping and identification of gene products from the lip-dacA-rodA-pbpA-leuS region of the Escherichia coli chromosome. J Bacteriol. 1980 Aug;143(2):569–581. doi: 10.1128/jb.143.2.569-581.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Spratt B. G. Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2999–3003. doi: 10.1073/pnas.72.8.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Spratt B. G. Escherichia coli resistance to beta-lactam antibiotics through a decrease in the affinity of a target for lethality. Nature. 1978 Aug 17;274(5672):713–715. doi: 10.1038/274713a0. [DOI] [PubMed] [Google Scholar]
  39. Spratt B. G., Pardee A. B. Penicillin-binding proteins and cell shape in E. coli. Nature. 1975 Apr 10;254(5500):516–517. doi: 10.1038/254516a0. [DOI] [PubMed] [Google Scholar]
  40. Spratt B. G. The mechanism of action of mecillinam. J Antimicrob Chemother. 1977 Jul;3 (Suppl B):13–19. doi: 10.1093/jac/3.suppl_b.13. [DOI] [PubMed] [Google Scholar]
  41. Tamaki S., Matsuzawa H., Matsuhashi M. Cluster of mrdA and mrdB genes responsible for the rod shape and mecillinam sensitivity of Escherichia coli. J Bacteriol. 1980 Jan;141(1):52–57. doi: 10.1128/jb.141.1.52-57.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tucker S. D., Gopalakrishnan A. S., Bollinger R., Dowhan W., Murgola E. J. Molecular mapping of glyW, a duplicate gene for tRNA3Gly of Escherichia coli. J Bacteriol. 1982 Nov;152(2):773–779. doi: 10.1128/jb.152.2.773-779.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wachi M., Doi M., Tamaki S., Park W., Nakajima-Iijima S., Matsuhashi M. Mutant isolation and molecular cloning of mre genes, which determine cell shape, sensitivity to mecillinam, and amount of penicillin-binding proteins in Escherichia coli. J Bacteriol. 1987 Nov;169(11):4935–4940. doi: 10.1128/jb.169.11.4935-4940.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Waxman D. J., Strominger J. L. Penicillin-binding proteins and the mechanism of action of beta-lactam antibiotics. Annu Rev Biochem. 1983;52:825–869. doi: 10.1146/annurev.bi.52.070183.004141. [DOI] [PubMed] [Google Scholar]
  45. Westling-Häggström B., Normark S. Genetic and physiological analysis of an envB spherelike mutant of Escherichia coli K-12 and characterization of its transductants. J Bacteriol. 1975 Jul;123(1):75–82. doi: 10.1128/jb.123.1.75-82.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Woldringh C. L., Binnerts J. S., Mans A. Variation in Escherichia coli buoyant density measured in Percoll gradients. J Bacteriol. 1981 Oct;148(1):58–63. doi: 10.1128/jb.148.1.58-63.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES