Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Apr;146(1):251–259. doi: 10.1128/jb.146.1.251-259.1981

Location of an F-pilin pool in the inner membrane.

D Moore, B A Sowa, K Ippen-Ihler
PMCID: PMC217076  PMID: 6111549

Abstract

Polyacrylamide gel analysis of [35S]methionine-labeled membrane preparations from Escherichia coli has revealed the presence of five polypeptides present only in the membranes of cells containing the conjugative plasmid F. In addition to the previously reported product of traT, polypeptides migrating with apparent molecular weights of 100,000, 23,500, 12,000, and 7,000 were resolved. Membrane preparations from F traJ mutants lacked these polypeptides, indicating that all of these proteins are tra gene products. The 7,000-molecular-weight polypeptide comigrated with unlabeled purified F-pilin protein. About 4 to 5% of the total radioactive label in whole membrane preparations was present in this polypeptide, indicating the existence of a substantial pool of membrane-associated F-pilin. The polypeptide could be extracted from whole membrane preparations with Triton X-100 and was found in the inner membrane fraction of membranes separated by sucrose density centrifugation.

Full text

PDF
251

Images in this article

253Image
on p.253
255Image
on p.255
257Image
on p.257

Selected References

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

  1. Achtman M., Kennedy N., Skurray R. Cell--cell interactions in conjugating Escherichia coli: role of traT protein in surface exclusion. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5104–5108. doi: 10.1073/pnas.74.11.5104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Achtman M., Manning P. A., Edelbluth C., Herrlich P. Export without proteolytic processing of inner and outer membrane proteins encoded by F sex factor tra cistrons in Escherichia coli minicells. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4837–4841. doi: 10.1073/pnas.76.10.4837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Achtman M., Manning P. A., Kusecek B., Schwuchow S., Willetts N. A genetic analysis of F sex factor cistrons needed for surface exclusion in Escherichia coli. J Mol Biol. 1980 Apr 25;138(4):779–795. doi: 10.1016/0022-2836(80)90065-0. [DOI] [PubMed] [Google Scholar]
  4. Achtman M., Willetts N., Clark A. J. Beginning a genetic analysis of conjugational transfer determined by the F factor in Escherichia coli by isolation and characterization of transfer-deficient mutants. J Bacteriol. 1971 May;106(2):529–538. doi: 10.1128/jb.106.2.529-538.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Armstrong G. D., Frost L. S., Sastry P. A., Paranchych W. Comparative biochemical studies on F and EDP208 conjugative pili. J Bacteriol. 1980 Jan;141(1):333–341. doi: 10.1128/jb.141.1.333-341.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beard J. P., Connolly J. C. Detection of a protein, similar to the sex pilus subunit, in the outer membrane of Escherichia coli cells carrying a derepressed F-like R factor. J Bacteriol. 1975 Apr;122(1):59–65. doi: 10.1128/jb.122.1.59-65.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brinton C. C., Jr The properties of sex pili, the viral nature of "conjugal" genetic transfer systems, and some possible approaches to the control of bacterial drug resistance. CRC Crit Rev Microbiol. 1971 May;1(1):105–160. doi: 10.3109/10408417109104479. [DOI] [PubMed] [Google Scholar]
  8. Brinton C. C., Jr The structure, function, synthesis and genetic control of bacterial pili and a molecular model for DNA and RNA transport in gram negative bacteria. Trans N Y Acad Sci. 1965 Jun;27(8):1003–1054. doi: 10.1111/j.2164-0947.1965.tb02342.x. [DOI] [PubMed] [Google Scholar]
  9. Curtiss R., 3rd Bacterial conjugation. Annu Rev Microbiol. 1969;23:69–136. doi: 10.1146/annurev.mi.23.100169.000441. [DOI] [PubMed] [Google Scholar]
  10. Date T., Inuzuka M., Tomoeda M. Purification and characterization of F pili from Escherichia coli. Biochemistry. 1977 Dec 13;16(25):5579–5585. doi: 10.1021/bi00644a030. [DOI] [PubMed] [Google Scholar]
  11. Everett R., Willetts N. Characterisation of an in vivo system for nicking at the origin of conjugal DNA transfer of the sex factor F. J Mol Biol. 1980 Jan 15;136(2):129–150. doi: 10.1016/0022-2836(80)90309-5. [DOI] [PubMed] [Google Scholar]
  12. Helmuth R., Achtman M. Cell-cell interactions in conjugating Escherichia coli: purification of F pili with biological activity. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1237–1241. doi: 10.1073/pnas.75.3.1237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kennedy N., Beutin L., Achtman M., Skurray R., Rahmsdorf U., Herrlich P. Conjugation proteins encoded by the F sex factor. Nature. 1977 Dec 15;270(5638):580–585. doi: 10.1038/270580a0. [DOI] [PubMed] [Google Scholar]
  14. Lutkenhaus J. F., Moore B. A., Masters M., Donachie W. D. Individual proteins are synthesized continuously throughout the Escherichia coli cell cycle. J Bacteriol. 1979 May;138(2):352–360. doi: 10.1128/jb.138.2.352-360.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marvin D. A., Hohn B. Filamentous bacterial viruses. Bacteriol Rev. 1969 Jun;33(2):172–209. doi: 10.1128/br.33.2.172-209.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Minkley E. G., Jr, Ippen-Ihler K. Identification of a membrane protein associated with expression of the surface exclusion region of the F transfer operon. J Bacteriol. 1977 Mar;129(3):1613–1622. doi: 10.1128/jb.129.3.1613-1622.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Minkley E. G., Jr, Polen S., Brinton C. C., Jr, Ippen-Ihler K. Identification of the structural gene for F-pilin. J Mol Biol. 1976 Nov;108(1):111–121. doi: 10.1016/s0022-2836(76)80098-8. [DOI] [PubMed] [Google Scholar]
  18. Novotny C. P., Fives-Taylor P. Retraction of F pili. J Bacteriol. 1974 Mar;117(3):1306–1311. doi: 10.1128/jb.117.3.1306-1311.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  20. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  21. Ridgway H. G., Silverman M., Simon M. I. Localization of proteins controlling motility and chemotaxis in Escherichia coli. J Bacteriol. 1977 Nov;132(2):657–665. doi: 10.1128/jb.132.2.657-665.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  23. Willetts N. S., Moore P. M., Paranchych W. Variant pili produced by mutants of the Flac plasmid. J Gen Microbiol. 1980 Apr;117(2):455–464. doi: 10.1099/00221287-117-2-455. [DOI] [PubMed] [Google Scholar]
  24. Willetts N., Maule J. Investigations of the F conjugation gene traI:traI mutants and lambdatraI transducing phages. Mol Gen Genet. 1979 Feb 1;169(3):325–336. doi: 10.1007/BF00382278. [DOI] [PubMed] [Google Scholar]
  25. Willetts N. The transcriptional control of fertility in F-like plasmids. J Mol Biol. 1977 May 5;112(1):141–148. doi: 10.1016/s0022-2836(77)80161-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES