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. 1988 Nov;170(11):5169–5176. doi: 10.1128/jb.170.11.5169-5176.1988

Identification, cloning, and expression of bolA, an ftsZ-dependent morphogene of Escherichia coli.

M Aldea 1, C Hernández-Chico 1, A G de la Campa 1, S R Kushner 1, M Vicente 1
PMCID: PMC211586  PMID: 3053647

Abstract

A newly found morphogene of Escherichia coli, bolA, mapping at min 10 of the genetic map, was cloned in a 7.2-kilobase BamHI fragment and identified by its ability to produce osmotically stable spherical cells when overexpressed. This gene codes for a polypeptide of 13 kilodaltons. Overexpression of bolA+ was achieved in low-copy-number vectors with operon fusions to the tet and lac promoters, indicating a clockwise direction of transcription. While no modification of any of the penicillin-binding proteins was observed, morphological effects due to overexpression of bolA+ were shown to be dependent on the presence of an active ftsZ gene product. Our results suggest the existence of a mechanism mediated by FtsZ for modifying the conformation of nascent murein in the early steps of septum formation.

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Selected References

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

  1. 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]
  2. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Begg K. J., Hatfull G. F., Donachie W. D. Identification of new genes in a cell envelope-cell division gene cluster of Escherichia coli: cell division gene ftsQ. J Bacteriol. 1980 Oct;144(1):435–437. doi: 10.1128/jb.144.1.435-437.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Begg K. J., Spratt B. G., Donachie W. D. Interaction between membrane proteins PBP3 and rodA is required for normal cell shape and division in Escherichia coli. J Bacteriol. 1986 Sep;167(3):1004–1008. doi: 10.1128/jb.167.3.1004-1008.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Berg C. M., Curtiss R., 3rd Transposition derivatives of an Hfr strain of Escherichia coli K-12. Genetics. 1967 Jul;56(3):503–525. doi: 10.1093/genetics/56.3.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Broome-Smith J. K., Spratt B. G. Deletion of the penicillin-binding protein 6 gene of Escherichia coli. J Bacteriol. 1982 Nov;152(2):904–906. doi: 10.1128/jb.152.2.904-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cesareni G., Cornelissen M., Lacatena R. M., Castagnoli L. Control of pMB1 replication: inhibition of primer formation by Rop requires RNA1. EMBO J. 1984 Jun;3(6):1365–1369. doi: 10.1002/j.1460-2075.1984.tb01978.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davis R., Vapnek D. In vivo transcription of R-plasmid deoxyribonucleic acid in Escherichia coli strains with altered antibiotic resistance levels and/or conjugal proficiency. J Bacteriol. 1976 Mar;125(3):1148–1155. doi: 10.1128/jb.125.3.1148-1155.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Donachie W. D., Begg K. J., Vicente M. Cell length, cell growth and cell division. Nature. 1976 Nov 25;264(5584):328–333. doi: 10.1038/264328a0. [DOI] [PubMed] [Google Scholar]
  12. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  13. Higgins M. L., Shockman G. D. Procaryotic cell division with respect to wall and membranes. CRC Crit Rev Microbiol. 1971 May;1(1):29–72. doi: 10.3109/10408417109104477. [DOI] [PubMed] [Google Scholar]
  14. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  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. 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]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Langridge J., Langridge P., Bergquist P. L. Extraction of nucleic acids from agarose gels. Anal Biochem. 1980 Apr;103(2):264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  20. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lutkenhaus J. F. Coupling of DNA replication and cell division: sulB is an allele of ftsZ. J Bacteriol. 1983 Jun;154(3):1339–1346. doi: 10.1128/jb.154.3.1339-1346.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Markiewicz Z., Broome-Smith J. K., Schwarz U., Spratt B. G. Spherical E. coli due to elevated levels of D-alanine carboxypeptidase. Nature. 1982 Jun 24;297(5868):702–704. doi: 10.1038/297702a0. [DOI] [PubMed] [Google Scholar]
  24. Martínez-Salas E., Vicente M. Amber mutation affecting the length of Escherichia coli cells. J Bacteriol. 1980 Nov;144(2):532–541. doi: 10.1128/jb.144.2.532-541.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mirelman D., Yashouv-Gan Y., Nuchamovitz Y., Rozenhak S., Ron E. Z. Murein biosynthesis during a synchromous cell cycle of Escherichia coli B. J Bacteriol. 1978 May;134(2):458–461. doi: 10.1128/jb.134.2.458-461.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mirelman D., Yashouv-Gan Y., Schwarz U. Regulation of murein biosynthesis and septum formation in filamentous cells of Escherichia coli PAT 84. J Bacteriol. 1977 Mar;129(3):1593–1600. doi: 10.1128/jb.129.3.1593-1600.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nishimura Y., Takeda Y., Nishimura A., Suzuki H., Inouye M., Hirota Y. Synthetic ColE1 plasmids carrying genes for cell division in Escherichia coli. Plasmid. 1977 Nov;1(1):67–77. doi: 10.1016/0147-619x(77)90009-9. [DOI] [PubMed] [Google Scholar]
  29. Pisabarro A. G., Prats R., Váquez D., Rodríguez-Tébar A. Activity of penicillin-binding protein 3 from Escherichia coli. J Bacteriol. 1986 Oct;168(1):199–206. doi: 10.1128/jb.168.1.199-206.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rogers H. J. Biogenesis of the wall in bacterial morphogenesis. Adv Microb Physiol. 1979;19:1–62. doi: 10.1016/s0065-2911(08)60197-6. [DOI] [PubMed] [Google Scholar]
  31. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. Stoker N. G., Fairweather N. F., Spratt B. G. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. doi: 10.1016/0378-1119(82)90172-x. [DOI] [PubMed] [Google Scholar]
  36. Taschner P. E., Huls P. G., Pas E., Woldringh C. L. Division behavior and shape changes in isogenic ftsZ, ftsQ, ftsA, pbpB, and ftsE cell division mutants of Escherichia coli during temperature shift experiments. J Bacteriol. 1988 Apr;170(4):1533–1540. doi: 10.1128/jb.170.4.1533-1540.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tormo A., Dopazo A., de la Campa A. G., Aldea M., Vicente M. Coupling between DNA replication and cell division mediated by the FtsA protein in Escherichia coli: a pathway independent of the SOS response, the "TER" pathway. J Bacteriol. 1985 Nov;164(2):950–953. doi: 10.1128/jb.164.2.950-953.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ward J. E., Jr, Lutkenhaus J. Overproduction of FtsZ induces minicell formation in E. coli. Cell. 1985 Oct;42(3):941–949. doi: 10.1016/0092-8674(85)90290-9. [DOI] [PubMed] [Google Scholar]
  39. de la Campa A. G., Martínez-Salas E., Tormo A., Vicente M. Co-ordination between elongation and division in Escherichia coli mediated by the wee gene product. J Gen Microbiol. 1984 Oct;130(10):2671–2679. doi: 10.1099/00221287-130-10-2671. [DOI] [PubMed] [Google Scholar]

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