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. 1993 Mar;175(5):1384–1391. doi: 10.1128/jb.175.5.1384-1391.1993

Synthesis of F pilin.

K Maneewannakul 1, S Maneewannakul 1, K Ippen-Ihler 1
PMCID: PMC193225  PMID: 8095257

Abstract

Transfer of the Escherichia coli fertility plasmid, F, is dependent on expression of F pili. Synthesis of F-pilin subunits is known to involve three F plasmid transfer (tra) region products: traA encodes the 13-kDa precursor protein, TraQ permits this to be processed to the 7-kDa pilin polypeptide, and TraX catalyzes acetylation of the pilin amino terminus. Using cloned tra sequences, we performed a series of pulse-chase experiments to investigate the effect of TraQ and TraX on the fate of the traA product. In TraQ- cells, the traA gene product was found to be very unstable. While traA polypeptides of various sizes were detected early in the chase period, almost all were degraded within 5 min. Rapid traA product degradation was also observed in TraX+ cells, although an increased percentage of these products persisted during the chase. In TraQ+ cells, most of the traA product was processed to the 7-kDa pilin polypeptide within the 1-min pulse period; this product [7(Q)] was not degraded but was increasingly converted to an 8-kDa form [8(Q)] as the chase continued, suggesting that host enzymes can modify the pilin polypeptide. Similar results were observed in TraQ+ TraX+ cells, but the primary 7-kDa product appeared to be N-acetylated pilin (Ac-7). An 8-kDa product (Ac-8) was also detected, but this band did not increase in intensity during the chase. We suggest a pathway in which TraQ prevents the traA product from folding to a readily degradable conformation and assists its entry into the membrane, Leader peptidase I cleaves the traA product signal sequence, and a subset of the pilin polypeptides becomes modified by host enzymes; TraX then acetylates the N terminal of both the modified and unmodified pilin polypeptides.

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Selected References

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

  1. AMBLER R. P., REES M. W. Epsilon-N-Methyl-lysine in bacterial flagellar protein. Nature. 1959 Jul 4;184:56–57. doi: 10.1038/184056b0. [DOI] [PubMed] [Google Scholar]
  2. Armstrong G. D., Frost L. S., Vogel H. J., Paranchych W. Nature of the carbohydrate and phosphate associated with ColB2 and EDP208 pilin. J Bacteriol. 1981 Mar;145(3):1167–1176. doi: 10.1128/jb.145.3.1167-1176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Cram D. S., Loh S. M., Cheah K. C., Skurray R. A. Sequence and conservation of genes at the distal end of the transfer region on plasmids F and R6-5. Gene. 1991 Jul 31;104(1):85–90. doi: 10.1016/0378-1119(91)90469-r. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Frost L. S., Bazett-Jones D. P. Examination of the phosphate in conjugative F-like pili by use of electron spectroscopic imaging. J Bacteriol. 1991 Dec;173(23):7728–7731. doi: 10.1128/jb.173.23.7728-7731.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Frost L. S., Lee J. S., Scraba D. G., Paranchych W. Two monoclonal antibodies specific for different epitopes within the amino-terminal region of F pilin. J Bacteriol. 1986 Oct;168(1):192–198. doi: 10.1128/jb.168.1.192-198.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frost L. S., Paranchych W., Willetts N. S. DNA sequence of the F traALE region that includes the gene for F pilin. J Bacteriol. 1984 Oct;160(1):395–401. doi: 10.1128/jb.160.1.395-401.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grossman T. H., Frost L. S., Silverman P. M. Structure and function of conjugative pili: monoclonal antibodies as probes for structural variants of F pili. J Bacteriol. 1990 Mar;172(3):1174–1179. doi: 10.1128/jb.172.3.1174-1179.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inokuchi K., Itoh M., Mizushima S. Domains involved in osmoregulation of the ompF gene in Escherichia coli. J Bacteriol. 1985 Nov;164(2):585–590. doi: 10.1128/jb.164.2.585-590.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ippen-Ihler K., Moore D., Laine S., Johnson D. A., Willetts N. S. Synthesis of F-pilin polypeptide in the absence of F traJ product. Plasmid. 1984 Mar;11(2):116–129. doi: 10.1016/0147-619x(84)90017-9. [DOI] [PubMed] [Google Scholar]
  12. Kathir P., Ippen-Ihler K. Construction and characterization of derivatives carrying insertion mutations in F plasmid transfer region genes, trbA, artA, traQ, and trbB. Plasmid. 1991 Jul;26(1):40–54. doi: 10.1016/0147-619x(91)90035-u. [DOI] [PubMed] [Google Scholar]
  13. Laine S., Moore D., Kathir P., Ippen-Ihler K. Genes and gene products involved in the synthesis of F-pili. Basic Life Sci. 1985;30:535–553. doi: 10.1007/978-1-4613-2447-8_38. [DOI] [PubMed] [Google Scholar]
  14. Maneewannakul K., Maneewannakul S., Ippen-Ihler K. Sequence alterations affecting F plasmid transfer gene expression: a conjugation system dependent on transcription by the RNA polymerase of phage T7. Mol Microbiol. 1992 Oct;6(20):2961–2973. doi: 10.1111/j.1365-2958.1992.tb01755.x. [DOI] [PubMed] [Google Scholar]
  15. Maneewannakul S., Kathir P., Ippen-Ihler K. Characterization of the F plasmid mating aggregation gene traN and of a new F transfer region locus trbE. J Mol Biol. 1992 May 20;225(2):299–311. doi: 10.1016/0022-2836(92)90923-8. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Moore D., Hamilton C. M., Maneewannakul K., Mintz Y., Frost L. S., Ippen-Ihler K. The Escherichia coli K-12 F plasmid gene traX is required for acetylation of F pilin. J Bacteriol. 1993 Mar;175(5):1375–1383. doi: 10.1128/jb.175.5.1375-1383.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moore D., Sowa B. A., Ippen-Ihler K. A new activity in the Ftra operon which is required for F-pilin synthesis. Mol Gen Genet. 1982;188(3):459–464. doi: 10.1007/BF00330049. [DOI] [PubMed] [Google Scholar]
  19. Moore D., Sowa B. A., Ippen-Ihler K. Location of an F-pilin pool in the inner membrane. J Bacteriol. 1981 Apr;146(1):251–259. doi: 10.1128/jb.146.1.251-259.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moore D., Sowa B. A., Ippen-Ihler K. The effect of tra mutations on the synthesis of the F-pilin membrane polypeptide. Mol Gen Genet. 1981;184(2):260–264. doi: 10.1007/BF00272914. [DOI] [PubMed] [Google Scholar]
  21. Moore D., Wu J. H., Kathir P., Hamilton C. M., Ippen-Ihler K. Analysis of transfer genes and gene products within the traB-traC region of the Escherichia coli fertility factor, F. J Bacteriol. 1987 Sep;169(9):3994–4002. doi: 10.1128/jb.169.9.3994-4002.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nilsson B., Anderson S. Proper and improper folding of proteins in the cellular environment. Annu Rev Microbiol. 1991;45:607–635. doi: 10.1146/annurev.mi.45.100191.003135. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Sowa B. A., Moore D., Ippen-Ihler K. Physiology of F-pilin synthesis and utilization. J Bacteriol. 1983 Feb;153(2):962–968. doi: 10.1128/jb.153.2.962-968.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weinstein B., Solomon F. Microtubule assembly and phage morphogenesis: new results and classical paradigms. Mol Microbiol. 1992 Mar;6(6):677–681. doi: 10.1111/j.1365-2958.1992.tb01515.x. [DOI] [PubMed] [Google Scholar]
  26. Wu J. H., Ippen-Ihler K. Nucleotide sequence of traQ and adjacent loci in the Escherichia coli K-12 F-plasmid transfer operon. J Bacteriol. 1989 Jan;171(1):213–221. doi: 10.1128/jb.171.1.213-221.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wu J. H., Moore D., Lee T., Ippen-Ihler K. Analysis of Escherichia coli K12 F factor transfer genes: traQ, trbA, and trbB. Plasmid. 1987 Jul;18(1):54–69. doi: 10.1016/0147-619x(87)90078-3. [DOI] [PubMed] [Google Scholar]

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