DOI: 10.1111/j.1365-2958.1995.tb02420.x. Characterization of the <i>pilF—pilD</i> pilus‐assembly locus of <i>Neisseria gonorrhoeae</i>

Characterization of the pilF—pilD pilus‐assembly locus of Neisseria gonorrhoeae

10.1111/j.1365-2958.1995.tb02420.x

Crossref journal-article

Wiley

Molecular Microbiology (311)

Abstract

SummaryExpression of Type IV pili by the bacterial pathogen Neisseria gonorrhoeae appears to be essential for colonization of the human host. Several N. gonorrhoeae gene products have been recently identified which bear homology to proteins involved in pilus assembly and protein export in other bacterial systems. We report here the isolation and characterization of transposon insertion mutants in N. gonorrhoeae whose phenotypes indicate that the N. gonorrhoeae pilF and pilD gene products are required for gonoccocal pilus biogenesis. Mutants lacking the pilD gene product, a pre‐pilin peptidase, were unable to process the pre‐pilin subunit into pilin and thus were non‐piliated. pilF mutants processed pilin but did not assemble the mature subunit. Both classes of mutants released S‐pilin, a soluble, truncated form of the pilin subunit previously correlated with defects in pilus assembly. In addition, mutants containing transposon insertions in pilD or in a downstream gene, orfX, exhibited a severely restricted growth phenotype. Deletion analysis of pilD indicated that the poor growth phenotype observed for the pilD transposon mutants was a result of polar effects of the insertions on orfX expression. orfX encodes a predicted polypeptide of 23 kDa which contains a consensus nucleotide‐binding domain and has apparent homologues in Pseudomonas aeruginosa, Pseudomonas putida, Thermus thermophilus, and the eukaryote Caenorhabditis elegans. Although expression of orfX and pilD appears to be transcriptionally coupled, mutants containing transposon insertions in orfX expressed pili. Unlike either pilF or pilD mutants, orfX mutants were also competent for DNA transformation.

Bibliography

Freitag, N. E., Seifert, H. S., & Koomey, M. (1995). Characterization of the pilFâpilD pilusâassembly locus of Neisseria gonorrhoeae. Molecular Microbiology, 16(3), 575â586. Portico.

Authors 3

Nancy E. Freitag (first)
H. Steven Seifert (additional)
Michael Koomey (additional)

References 56 Referenced 140

10.1128/jb.171.10.5386-5404.1989
10.1016/0147-619X(90)90020-D
{'key': 'e_1_2_1_4_1', 'volume-title': 'Current Protocols in Molecular Biology', 'author': 'Ausubel F.M.', 'year': '1991'} / Current Protocols in Molecular Biology by Ausubel F.M. (1991)
10.1128/jb.173.2.479-486.1991
10.1128/jb.129.2.983-992.1977 / J Bacteriol / Factors affecting genetic transformation of Neisseria gonorrhoeae by Biswas G.D. (1977)
{'key': 'e_1_2_1_7_1', 'first-page': 'S24', 'article-title': 'Gene transfer in Neisseria gonorrhoeae', 'volume': '2', 'author': 'Biswas G.D.', 'year': '1989', 'journal-title': 'J Bacteriol'} / J Bacteriol / Gene transfer in Neisseria gonorrhoeae by Biswas G.D. (1989)
10.1093/infdis/128.2.178
10.1111/j.1365-2958.1995.tb02267.x
10.1093/nar/12.1Part1.387
10.1128/MMBR.55.3.395-424.1991
10.1007/BF00267456
10.1128/jb.176.5.1323-1331.1994 / J Bacteriol / Type IV prepilin peptidase gene of Neisseria gonorrhoeae MS11: presence of a related gene in other pillated and nonpiliated Neisseria strains by Dupuy B. (1994)
10.1128/JB.160.3.1184-1187.1984
10.1016/0092-8674(86)90730-0
10.1126/science.1683004
10.1146/annurev.bi.57.070188.001215
10.1128/jb.176.3.642-650.1994 / J Bacteriol / Characterization of Type IV pilus genes in plant growth‐promoting Pseudomonas putida WCS358 by Groot A. (1994)
10.1073/pnas.84.24.9079
10.1128/CMR.2.Suppl.S66
10.1111/j.1365-2958.1993.tb01949.x
10.1002/j.1460-2075.1991.tb07970.x
10.1101/gad.5.10.1834
10.1128/jb.96.3.596-605.1968 / J Bacteriol / Neisseria gonorrhoeae II. Colonial variation and pathogenicity during 35 months in vitro by Kellog D.S. (1968)
10.1128/IAI.61.4.1371-1377.1993
10.1111/j.1365-2958.1991.tb02108.x
10.1093/genetics/117.3.391
10.1111/j.1365-2958.1993.tb01579.x
10.1128/JB.163.1.132-139.1985
{'key': 'e_1_2_1_30_1', 'first-page': '3165', 'article-title': 'On the role of pili in transformation of Neisseria gonorrboeae', 'volume': '130', 'author': 'Mathis L.S.', 'year': '1984', 'journal-title': 'J Gen Miorobiol'} / J Gen Miorobiol / On the role of pili in transformation of Neisseria gonorrboeae by Mathis L.S. (1984)
10.1093/infdis/143.3.413
10.1128/jb.171.11.6043-6051.1989
10.1128/jb.172.6.2911-2919.1990
10.1111/j.1365-2958.1994.tb01017.x
10.1111/j.1365-2958.1988.tb00073.x
10.1111/j.1742-4658.2006.05480.x
10.1146/annurev.ge.24.120190.000435
10.1111/j.1365-2958.1990.tb00604.x
10.1111/j.1365-2958.1992.tb02211.x
10.1016/0378-1119(93)90543-C
10.1128/jb.172.1.40-46.1990
10.1128/iai.60.3.1232-1236.1992 / Infect Immun / Characterization of a cryptic gene pair from Neisseria gonorrhoeae that is common to pathogenic Neisseria species by Seifert H.S. (1992)
10.1128/IAI.58.9.3042-3049.1990
10.1128/JB.92.5.1364-1371.1966
{'key': 'e_1_2_1_45_1', 'first-page': '155', 'volume-title': 'The Gonococcus', 'author': 'Sparling P.F.', 'year': '1977'} / The Gonococcus by Sparling P.F. (1977)
10.1128/jb.165.2.367-372.1986
10.1146/annurev.mi.47.100193.003025
10.1073/pnas.90.6.2404
10.1084/jem.137.3.571
10.1016/0092-8674(86)90449-6
10.1084/jem.165.5.1344
10.1099/00221287-137-10-2483
10.1111/j.1365-2958.1995.tb02410.x
10.1128/jb.175.16.4962-4969.1993
10.1002/j.1460-2075.1982.tb01276.x
10.1038/368032a0
10.1073/pnas.89.12.5366

Dates

Type	When
Created	18 years, 10 months ago (Oct. 27, 2006, 11:17 p.m.)
Deposited	1 year, 10 months ago (Oct. 26, 2023, 3:44 a.m.)
Indexed	22 hours, 49 minutes ago (Aug. 29, 2025, 6:07 a.m.)
Issued	30 years, 3 months ago (May 1, 1995)
Published	30 years, 3 months ago (May 1, 1995)
Published Online	18 years, 10 months ago (Oct. 27, 2006)
Published Print	30 years, 3 months ago (May 1, 1995)

Funders 0

None

BibTeX

@article{Freitag_1995, title={Characterization of the pilF—pilD pilus‐assembly locus of Neisseria gonorrhoeae}, volume={16}, ISSN={1365-2958}, url={http://dx.doi.org/10.1111/j.1365-2958.1995.tb02420.x}, DOI={10.1111/j.1365-2958.1995.tb02420.x}, number={3}, journal={Molecular Microbiology}, publisher={Wiley}, author={Freitag, Nancy E. and Seifert, H. Steven and Koomey, Michael}, year={1995}, month=may, pages={575–586} }

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