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. 1997 Nov;63(11):4150–4157. doi: 10.1128/aem.63.11.4150-4157.1997

A novel competence gene, comP, is essential for natural transformation of Acinetobacter sp. strain BD413.

D Porstendörfer 1, U Drotschmann 1, B Averhoff 1
PMCID: PMC168731  PMID: 9361398

Abstract

Acinetobacter sp. strain BD413 (= ATCC 33305), a nutritionally versatile bacterium, has an extremely efficient natural transformation system. Here we describe the generation of eight transformation-affected mutants of Acinetobacter sp. strain BD413 by insertional mutagenesis. These mutants were found by Southern blot analysis and complementation studies to result from single nptII marker insertions at different chromosomal loci. DNA binding and uptake studies with one mutant, T205, revealed that the transformation deficiency of this mutant results from a complete lack of DNA binding and, therefore, uptake activity. A novel competence gene essential for natural transformation, named comP, was cloned by complementation of mutant T205. The nucleotide sequence of comP was determined, and its deduced 15-kDa polypeptide displays significant similarities to type IV pilins. Analysis of the ultrastructure of a transformation-deficient comP mutant and the transformation-competent wild-type strain revealed that both are covered with bundle-forming thin fimbriae (3 to 4 nm in diameter) and individual thick fimbriae (6 nm in diameter). These results provide evidence that the pilinlike ComP is unrelated to the piluslike structures of strain BD413. Taking all data into account, we propose that ComP functions as a major subunit of an organelle acting as a channel or pore mediating DNA binding and/or uptake in Acinetobacter sp. strain BD413.

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Selected References

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  1. Albano M., Breitling R., Dubnau D. A. Nucleotide sequence and genetic organization of the Bacillus subtilis comG operon. J Bacteriol. 1989 Oct;171(10):5386–5404. doi: 10.1128/jb.171.10.5386-5404.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albano M., Dubnau D. A. Cloning and characterization of a cluster of linked Bacillus subtilis late competence mutations. J Bacteriol. 1989 Oct;171(10):5376–5385. doi: 10.1128/jb.171.10.5376-5385.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ausubel F. M. Regulation of nitrogen fixation genes. Cell. 1984 May;37(1):5–6. doi: 10.1016/0092-8674(84)90294-0. [DOI] [PubMed] [Google Scholar]
  4. Averhoff B., Gregg-Jolly L., Elsemore D., Ornston L. N. Genetic analysis of supraoperonic clustering by use of natural transformation in Acinetobacter calcoaceticus. J Bacteriol. 1992 Jan;174(1):200–204. doi: 10.1128/jb.174.1.200-204.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breitling R., Dubnau D. A membrane protein with similarity to N-methylphenylalanine pilins is essential for DNA binding by competent Bacillus subtilis. J Bacteriol. 1990 Mar;172(3):1499–1508. doi: 10.1128/jb.172.3.1499-1508.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doten R. C., Ngai K. L., Mitchell D. J., Ornston L. N. Cloning and genetic organization of the pca gene cluster from Acinetobacter calcoaceticus. J Bacteriol. 1987 Jul;169(7):3168–3174. doi: 10.1128/jb.169.7.3168-3174.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dubnau D. Genetic competence in Bacillus subtilis. Microbiol Rev. 1991 Sep;55(3):395–424. doi: 10.1128/mr.55.3.395-424.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Facius D., Fussenegger M., Meyer T. F. Sequential action of factors involved in natural competence for transformation of Neisseria gonorrhoeae. FEMS Microbiol Lett. 1996 Apr 1;137(2-3):159–164. doi: 10.1111/j.1574-6968.1996.tb08099.x. [DOI] [PubMed] [Google Scholar]
  10. Gibbs C. P., Reimann B. Y., Schultz E., Kaufmann A., Haas R., Meyer T. F. Reassortment of pilin genes in Neisseria gonorrhoeae occurs by two distinct mechanisms. Nature. 1989 Apr 20;338(6217):651–652. doi: 10.1038/338651a0. [DOI] [PubMed] [Google Scholar]
  11. Goodgal S. H. DNA uptake in Haemophilus transformation. Annu Rev Genet. 1982;16:169–192. doi: 10.1146/annurev.ge.16.120182.001125. [DOI] [PubMed] [Google Scholar]
  12. Gregg-Jolly L. A., Ornston L. N. Properties of Acinetobacter calcoaceticus recA and its contribution to intracellular gene conversion. Mol Microbiol. 1994 Jun;12(6):985–992. doi: 10.1111/j.1365-2958.1994.tb01086.x. [DOI] [PubMed] [Google Scholar]
  13. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  14. Hartnett G. B., Averhoff B., Ornston L. N. Selection of Acinetobacter calcoaceticus mutants deficient in the p-hydroxybenzoate hydroxylase gene (pobA), a member of a supraoperonic cluster. J Bacteriol. 1990 Oct;172(10):6160–6161. doi: 10.1128/jb.172.10.6160-6161.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Henrichsen J., Blom J. Correlation between twitching motility and possession of polar fimbriae in Acinetobacter calcoaceticus. Acta Pathol Microbiol Scand B. 1975 Apr;83(2):103–115. doi: 10.1111/j.1699-0463.1975.tb00078.x. [DOI] [PubMed] [Google Scholar]
  16. Henrichsen J. Twitching motility. Annu Rev Microbiol. 1983;37:81–93. doi: 10.1146/annurev.mi.37.100183.000501. [DOI] [PubMed] [Google Scholar]
  17. Hobbs M., Mattick J. S. Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes. Mol Microbiol. 1993 Oct;10(2):233–243. doi: 10.1111/j.1365-2958.1993.tb01949.x. [DOI] [PubMed] [Google Scholar]
  18. Inoue H., Nojima H., Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene. 1990 Nov 30;96(1):23–28. doi: 10.1016/0378-1119(90)90336-p. [DOI] [PubMed] [Google Scholar]
  19. Jonsson A. B., Nyberg G., Normark S. Phase variation of gonococcal pili by frameshift mutation in pilC, a novel gene for pilus assembly. EMBO J. 1991 Feb;10(2):477–488. doi: 10.1002/j.1460-2075.1991.tb07970.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jonsson A. B., Pfeifer J., Normark S. Neisseria gonorrhoeae PilC expression provides a selective mechanism for structural diversity of pili. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3204–3208. doi: 10.1073/pnas.89.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Juni E., Janik A. Transformation of Acinetobacter calco-aceticus (Bacterium anitratum). J Bacteriol. 1969 Apr;98(1):281–288. doi: 10.1128/jb.98.1.281-288.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. doi: 10.1016/0378-1119(88)90117-5. [DOI] [PubMed] [Google Scholar]
  23. Lorenz M. G., Reipschläger K., Wackernagel W. Plasmid transformation of naturally competent Acinetobacter calcoaceticus in non-sterile soil extract and groundwater. Arch Microbiol. 1992;157(4):355–360. doi: 10.1007/BF00248681. [DOI] [PubMed] [Google Scholar]
  24. Lorenz M. G., Wackernagel W. Bacterial gene transfer by natural genetic transformation in the environment. Microbiol Rev. 1994 Sep;58(3):563–602. doi: 10.1128/mr.58.3.563-602.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mazodier P., Davies J. Gene transfer between distantly related bacteria. Annu Rev Genet. 1991;25:147–171. doi: 10.1146/annurev.ge.25.120191.001051. [DOI] [PubMed] [Google Scholar]
  26. Palmen R., Vosman B., Buijsman P., Breek C. K., Hellingwerf K. J. Physiological characterization of natural transformation in Acinetobacter calcoaceticus. J Gen Microbiol. 1993 Feb;139(2):295–305. doi: 10.1099/00221287-139-2-295. [DOI] [PubMed] [Google Scholar]
  27. Palmen R., Vosman B., Kok R., van der Zee J. R., Hellingwerf K. J. Characterization of transformation-deficient mutants of Acinetobacter calcoaceticus. Mol Microbiol. 1992 Jul;6(13):1747–1754. doi: 10.1111/j.1365-2958.1992.tb01347.x. [DOI] [PubMed] [Google Scholar]
  28. Paranchych W., Frost L. S. The physiology and biochemistry of pili. Adv Microb Physiol. 1988;29:53–114. doi: 10.1016/s0065-2911(08)60346-x. [DOI] [PubMed] [Google Scholar]
  29. Parge H. E., Forest K. T., Hickey M. J., Christensen D. A., Getzoff E. D., Tainer J. A. Structure of the fibre-forming protein pilin at 2.6 A resolution. Nature. 1995 Nov 2;378(6552):32–38. doi: 10.1038/378032a0. [DOI] [PubMed] [Google Scholar]
  30. Pugsley A. P. Multimers of the precursor of a type IV pilin-like component of the general secretory pathway are unrelated to pili. Mol Microbiol. 1996 Jun;20(6):1235–1245. doi: 10.1111/j.1365-2958.1996.tb02643.x. [DOI] [PubMed] [Google Scholar]
  31. Rudel T., Facius D., Barten R., Scheuerpflug I., Nonnenmacher E., Meyer T. F. Role of pili and the phase-variable PilC protein in natural competence for transformation of Neisseria gonorrhoeae. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7986–7990. doi: 10.1073/pnas.92.17.7986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Seifert H. S., Ajioka R. S., Marchal C., Sparling P. F., So M. DNA transformation leads to pilin antigenic variation in Neisseria gonorrhoeae. Nature. 1988 Nov 24;336(6197):392–395. doi: 10.1038/336392a0. [DOI] [PubMed] [Google Scholar]
  34. Smith H. O., Danner D. B., Deich R. A. Genetic transformation. Annu Rev Biochem. 1981;50:41–68. doi: 10.1146/annurev.bi.50.070181.000353. [DOI] [PubMed] [Google Scholar]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Strom M. S., Lory S. Structure-function and biogenesis of the type IV pili. Annu Rev Microbiol. 1993;47:565–596. doi: 10.1146/annurev.mi.47.100193.003025. [DOI] [PubMed] [Google Scholar]
  37. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]

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