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. 1989 Apr;171(4):2142–2147. doi: 10.1128/jb.171.4.2142-2147.1989

Assembly of mutant pilins in Pseudomonas aeruginosa: formation of pili composed of heterologous subunits.

B L Pasloske 1, D G Scraba 1, W Paranchych 1
PMCID: PMC209869  PMID: 2564847

Abstract

Recently, we reported the degree of N-terminal processing within the cytoplasmic membranes of three mutant pilins from Pseudomonas aeruginosa PAK with respect to leader peptide removal and the methylation of the N-terminal phenylalanine (B. L. Pasloske and W. Paranchych, Mol. Microbiol. 2:489-495, 1988). The results of those experiments showed that the deletion of 4 or 8 amino acids within the highly conserved N terminus greatly inhibited leader peptide removal. On the other hand, the mutation of the glutamate at position 5 to a lysine permitted leader peptide cleavage but inhibited transmethylase activity. In this report, we have examined the effects of these mutant pilins upon pilus assembly in a P. aeruginosa PAO host with or without the chromosomally encoded pilin gene present. Pilins with deletions of 4 or 8 amino acids in the N-terminal region were not incorporated into pili. Interestingly, pilin subunits containing the glutamate-to-lysine mutation were incorporated into compound pili together with PAO wild-type subunits. However, the mutant pilins were unable to polymerize as a homopolymer. When wild-type PAK and PAO pilin subunits were expressed in the same bacterial strain, the pilin subunits assembled into homopolymeric pili containing one or the other type of subunit.

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Selected References

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

  1. Bodey G. P., Bolivar R., Fainstein V., Jadeja L. Infections caused by Pseudomonas aeruginosa. Rev Infect Dis. 1983 Mar-Apr;5(2):279–313. doi: 10.1093/clinids/5.2.279. [DOI] [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Bradley D. E., Pitt T. L. Pilus-dependence of four Pseudomonas aeruginosa bacteriophages with non-contractile tails. J Gen Virol. 1974 Jul;24(1):1–15. doi: 10.1099/0022-1317-24-1-1. [DOI] [PubMed] [Google Scholar]
  4. Bradley D. E. The adsorption of Pseudomonas aeruginosa pilus-dependent bacteriophages to a host mutant with nonretractile pili. Virology. 1974 Mar;58(1):149–163. doi: 10.1016/0042-6822(74)90150-0. [DOI] [PubMed] [Google Scholar]
  5. Dalrymple B., Mattick J. S. An analysis of the organization and evolution of type 4 fimbrial (MePhe) subunit proteins. J Mol Evol. 1987;25(3):261–269. doi: 10.1007/BF02100020. [DOI] [PubMed] [Google Scholar]
  6. Doig P., Todd T., Sastry P. A., Lee K. K., Hodges R. S., Paranchych W., Irvin R. T. Role of pili in adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells. Infect Immun. 1988 Jun;56(6):1641–1646. doi: 10.1128/iai.56.6.1641-1646.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elleman T. C., Peterson J. E. Expression of multiple types of N-methyl Phe pili in Pseudomonas aeruginosa. Mol Microbiol. 1987 Nov;1(3):377–380. doi: 10.1111/j.1365-2958.1987.tb01945.x. [DOI] [PubMed] [Google Scholar]
  8. Folkhard W., Marvin D. A., Watts T. H., Paranchych W. Structure of polar pili from Pseudomonas aeruginosa strains K and O. J Mol Biol. 1981 Jun 15;149(1):79–93. doi: 10.1016/0022-2836(81)90261-8. [DOI] [PubMed] [Google Scholar]
  9. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  10. Goldberg J. B., Ohman D. E. Cloning and expression in Pseudomonas aeruginosa of a gene involved in the production of alginate. J Bacteriol. 1984 Jun;158(3):1115–1121. doi: 10.1128/jb.158.3.1115-1121.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grasberger B., Minton A. P., DeLisi C., Metzger H. Interaction between proteins localized in membranes. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6258–6262. doi: 10.1073/pnas.83.17.6258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Henrichsen J. Twitching motility. Annu Rev Microbiol. 1983;37:81–93. doi: 10.1146/annurev.mi.37.100183.000501. [DOI] [PubMed] [Google Scholar]
  13. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  14. 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]
  15. Pasloske B. L., Carpenter M. R., Frost L. S., Finlay B. B., Paranchych W. The expression of Pseudomonas aeruginosa PAK pilin gene mutants in Escherichia coli. Mol Microbiol. 1988 Mar;2(2):185–195. doi: 10.1111/j.1365-2958.1988.tb00020.x. [DOI] [PubMed] [Google Scholar]
  16. Pasloske B. L., Finlay B. B., Paranchych W. Cloning and sequencing of the Pseudomonas aeruginosa PAK pilin gene. FEBS Lett. 1985 Apr 22;183(2):408–412. doi: 10.1016/0014-5793(85)80821-8. [DOI] [PubMed] [Google Scholar]
  17. Pasloske B. L., Paranchych W. The expression of mutant pilins in Pseudomonas aeruginosa: fifth position glutamate affects pilin methylation. Mol Microbiol. 1988 Jul;2(4):489–495. doi: 10.1111/j.1365-2958.1988.tb00055.x. [DOI] [PubMed] [Google Scholar]
  18. Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
  19. Sastry P. A., Finlay B. B., Pasloske B. L., Paranchych W., Pearlstone J. R., Smillie L. B. Comparative studies of the amino acid and nucleotide sequences of pilin derived from Pseudomonas aeruginosa PAK and PAO. J Bacteriol. 1985 Nov;164(2):571–577. doi: 10.1128/jb.164.2.571-577.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sato H., Okinaga K., Saito H. Role of pili in the pathogenesis of Pseudomonas aeruginosa burn infection. Microbiol Immunol. 1988;32(2):131–139. doi: 10.1111/j.1348-0421.1988.tb01372.x. [DOI] [PubMed] [Google Scholar]
  21. Watts T. H., Kay C. M., Paranchych W. Spectral properties of three quaternary arrangements of Pseudomonas pilin. Biochemistry. 1983 Jul 19;22(15):3640–3646. doi: 10.1021/bi00284a016. [DOI] [PubMed] [Google Scholar]
  22. Watts T. H., Worobec E. A., Paranchych W. Identification of pilin pools in the membranes of Pseudomonas aeruginosa. J Bacteriol. 1982 Nov;152(2):687–691. doi: 10.1128/jb.152.2.687-691.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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