The comP locus of Neisseria gonorrhoeae encodes a type IV prepilin that is dispensable for pilus biogenesis but essential for natural transformation

ABSTRACT The extreme thermophile Thermus thermophilus HB27 exhibits high frequencies of natural transformation. Although we recently reported identification of the first competence genes in Thermus, the molecular basis of DNA uptake is unknown. A pilus-like structure is assumed to be involved. Twelve genes encoding prepilin-like proteins were identified in three loci in the genome of T. thermophilus. Mutational analyses, described in this paper, revealed that one locus, which contains four genes that encode prepilin-like proteins (pilA1 to pilA4), is essential for natural transformation. Additionally, comZ, a new competence gene with no similarity to known genes, was identified. Analysis of the piliation phenotype revealed wild-type piliation of a pilA1-pilA3Δkat mutant and a comZ mutant, whereas a pilA4 mutant was found to be completely devoid of pilus structures. These findings, together with the significant similarity of PilA4 to prepilins, led to the conclusion that the T. thermophilus pilus structures are type IV pili. Furthermore, the loss of the transformation and piliation phenotype in the pilA4 mutant suggests that type IV pili are implicated in natural transformation of T. thermophilus HB27.

Download Full-text

DNA Uptake Sequence-Mediated Enhancement of Transformation in Neisseria gonorrhoeae Is Strain Dependent

Journal of Bacteriology ◽

10.1128/jb.00442-10 ◽

2010 ◽

Vol 192 (17) ◽

pp. 4436-4444 ◽

Cited By ~ 30

Author(s):

Paul M. Duffin ◽

H. Steven Seifert

Keyword(s):

Neisseria Gonorrhoeae ◽

Plasmid Dna ◽

Natural Transformation ◽

Dna Transformation ◽

Dna Uptake ◽

Efficient Transformation ◽

Type Iv ◽

Different Strains ◽

Strain Dependent

ABSTRACT Natural transformation is the main means of horizontal genetic exchange in the obligate human pathogen Neisseria gonorrhoeae. Neisseria spp. have been shown to preferentially take up and transform their own DNA by recognizing the nonpalindromic 10- or 12-nucleotide sequence 5′- AT GCCGTCTGAA-3′ (additional semiconserved nucleotides are underlined), termed the DNA uptake sequence (DUS10 or DUS12). Here we investigated the effects of the DUS on transformation and DNA uptake for several laboratory strains of N. gonorrhoeae. We found that all strains showed efficient transformation of DUS containing DNA (DUS10 and DUS12) but that the level of transformation with DNA lacking a DUS (DUS0) was variable in different strains. The DUS-enhanced transformation was 20-fold in two strains, FA1090 and FA19, but was approximately 150-fold in strains MS11 and 1291. All strains tested provide some level of DUS0 transformation, and DUS0 transformation was type IV pilus dependent. Competition with plasmid DNA revealed that transformation of MS11 was enhanced by the addition of excess plasmid DNA containing a DUS while FA1090 transformation was competitively inhibited. Although FA1090 was able to mediate much more efficient transformation of DNA lacking a DUS than was MS11, DNA uptake experiments showed similar levels of uptake of DNA containing and lacking a DUS in FA1090 and MS11. Finally, DNA uptake was competitively inhibited in both FA1090 and MS11. Taken together, our data indicate that the role of the DUS during DNA transformation is variable between strains of N. gonorrhoeae and may influence multiple steps during transformation.

Download Full-text

Competence for natural transformation in Neisseria gonorrhoeae: components of DNA binding and uptake linked to type IV pilus expression

Molecular Microbiology ◽

10.1046/j.1365-2958.2002.03193.x ◽

2002 ◽

Vol 46 (3) ◽

pp. 749-760 ◽

Cited By ~ 104

Author(s):

Finn Erik Aas ◽

Matthew Wolfgang ◽

Stephan Frye ◽

Steven Dunham ◽

Cecilia Løvold ◽

...

Keyword(s):

Dna Binding ◽

Neisseria Gonorrhoeae ◽

Natural Transformation ◽

Type Iv Pilus ◽

Type Iv

Download Full-text

Faculty Opinions recommendation of Natural transformation competence in Helicobacter pylori is mediated by the basic components of a type IV secretion system.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1003976.44256 ◽

2002 ◽

Author(s):

Christoph Dehio

Keyword(s):

Helicobacter Pylori ◽

Natural Transformation ◽

Secretion System ◽

Type Iv Secretion ◽

Type Iv Secretion System ◽

Type Iv ◽

Transformation Competence ◽

Natural Transformation Competence

Download Full-text

Type IV Pilus Assembly Proficiency and Dynamics Influence Pilin Subunit Phospho-Form Macro- and Microheterogeneity in Neisseria gonorrhoeae

PLoS ONE ◽

10.1371/journal.pone.0096419 ◽

2014 ◽

Vol 9 (5) ◽

pp. e96419 ◽

Cited By ~ 1

Author(s):

Åshild Vik ◽

Jan Haug Anonsen ◽

Finn Erik Aas ◽

Finn Terje Hegge ◽

Norbert Roos ◽

...

Keyword(s):

Neisseria Gonorrhoeae ◽

Type Iv Pilus ◽

Type Iv ◽

Pilin Subunit ◽

Pilus Assembly

Download Full-text

Identification and initial characterization of a new pair of sibling sRNAs of Neisseria gonorrhoeae involved in type IV pilus biogenesis

Microbiology ◽

10.1099/mic.0.001080 ◽

2021 ◽

Vol 167 (9) ◽

Author(s):

Marie Zachary ◽

Susanne Bauer ◽

Maximilian Klepsch ◽

Katharina Wagler ◽

Bruno Hüttel ◽

...

Keyword(s):

Target Genes ◽

Target Prediction ◽

Type Iv Pilus ◽

Content Type ◽

Type Iv ◽

Gene Expression Control ◽

Initial Characterization ◽

Pilus Biogenesis ◽

Regulatory Rnas

Non-coding regulatory RNAs mediate post-transcriptional gene expression control by a variety of mechanisms relying mostly on base-pairing interactions with a target mRNA. Though a plethora of putative non-coding regulatory RNAs have been identified by global transcriptome analysis, knowledge about riboregulation in the pathogenic Neisseriae is still limited. Here we report the initial characterization of a pair of sRNAs of N. gonorrhoeae , TfpR1 and TfpR2, which exhibit a similar secondary structure and identical single-stranded seed regions, and therefore might be considered as sibling sRNAs. By combination of in silico target prediction and sRNA pulse expression followed by differential RNA sequencing we identified target genes of TfpR1 which are involved in type IV pilus biogenesis and DNA damage repair. We provide evidence that members of the TfpR1 regulon can also be targeted by the sibling TfpR2.

Download Full-text

Purification and Three-Dimensional Electron Microscopy Structure of the Neisseria meningitidis Type IV Pilus Biogenesis Protein PilG

Journal of Bacteriology ◽

10.1128/jb.00648-07 ◽

2007 ◽

Vol 189 (17) ◽

pp. 6389-6396 ◽

Cited By ~ 25

Author(s):

Richard F. Collins ◽

Muhammad Saleem ◽

Jeremy P. Derrick

Keyword(s):

Electron Microscopy ◽

Neisseria Meningitidis ◽

Metal Ion ◽

Polyacrylamide Gel Electrophoresis ◽

Three Dimensional ◽

Type Ii Secretion ◽

Type Ii ◽

Type Iv Pilus ◽

Type Iv ◽

Pilus Biogenesis

ABSTRACT Type IV pili are surface-exposed retractable fibers which play a key role in the pathogenesis of Neisseria meningitidis and other gram-negative pathogens. PilG is an integral inner membrane protein and a component of the type IV pilus biogenesis system. It is related by sequence to the extensive GspF family of secretory proteins, which are involved in type II secretion processes. PilG was overexpressed and purified from Escherichia coli membranes by detergent extraction and metal ion affinity chromatography. Analysis of the purified protein by perfluoro-octanoic acid polyacrylamide gel electrophoresis showed that PilG formed dimers and tetramers. A three-dimensional (3-D) electron microscopy structure of the PilG multimer was determined using single-particle averaging applied to samples visualized by negative staining. Symmetry analysis of the unsymmetrized 3-D volume provided further evidence that the PilG multimer is a tetramer. The reconstruction also revealed an asymmetric bilobed structure approximately 125 Å in length and 80 Å in width. The larger lobe within the structure was identified as the N terminus by location of Ni-nitrilotriacetic acid nanogold particles to the N-terminal polyhistidine tag. We propose that the smaller lobe corresponds to the periplasmic domain of the protein, with the narrower “waist” region being the transmembrane section. This constitutes the first report of a 3-D structure of a member of the GspF family and suggests a physical basis for the role of the protein in linking cytoplasmic and periplasmic protein components of the type II secretion and type IV pilus biogenesis systems.

Download Full-text

The role of core and accessory type IV pilus genes in natural transformation and twitching motility in the bacterium Acinetobacter baylyi

PLoS ONE ◽

10.1371/journal.pone.0182139 ◽

2017 ◽

Vol 12 (8) ◽

pp. e0182139 ◽

Cited By ~ 18

Author(s):

Colleen G. Leong ◽

Rebecca A. Bloomfield ◽

Caroline A. Boyd ◽

Amber J. Dornbusch ◽

Leah Lieber ◽

...

Keyword(s):

Natural Transformation ◽

Twitching Motility ◽

Type Iv Pilus ◽

Acinetobacter Baylyi ◽

Type Iv

Download Full-text

Dynamics of Neisseria gonorrhoeae Attachment: Microcolony Development, Cortical Plaque Formation, and Cytoprotection

Infection and Immunity ◽

10.1128/iai.00687-07 ◽

2007 ◽

Vol 75 (10) ◽

pp. 4743-4753 ◽

Cited By ~ 69

Author(s):

Dustin L. Higashi ◽

Shaun W. Lee ◽

Aurelie Snyder ◽

Nathan J. Weyand ◽

Antony Bakke ◽

...

Keyword(s):

Neisseria Gonorrhoeae ◽

Transmitted Disease ◽

Virus Transmission ◽

Twitching Motility ◽

Sexually Transmitted ◽

Type Iv ◽

Immunodeficiency Virus ◽

Live Cell Microscopy ◽

Induced Apoptosis ◽

First Time

ABSTRACT Neisseria gonorrhoeae is the bacterium that causes gonorrhea, a major sexually transmitted disease and a significant cofactor for human immunodeficiency virus transmission. The retactile N. gonorrhoeae type IV pilus (Tfp) mediates twitching motility and attachment. Using live-cell microscopy, we reveal for the first time the dynamics of twitching motility by N. gonorrhoeae in its natural environment, human epithelial cells. Bacteria aggregate into microcolonies on the cell surface and induce a massive remodeling of the microvillus architecture. Surprisingly, the microcolonies are motile, and they fuse to form progressively larger structures that undergo rapid reorganization, suggesting that bacteria communicate with each other during infection. As reported, actin plaques form beneath microcolonies. Here, we show that cortical plaques comigrate with motile microcolonies. These activities are dependent on pilT, the Tfp retraction locus. Cultures infected with a pilT mutant have significantly higher numbers of apoptotic cells than cultures infected with the wild-type strain. Inducing pilT expression with isopropyl-β-d-thiogalactopyranoside partially rescues cells from infection-induced apoptosis, demonstrating that Tfp retraction is intrinsically cytoprotective for the host. Tfp-mediated attachment is therefore a continuum of microcolony motility and force stimulation of host cell signaling, leading to a cytoprotective effect.

Download Full-text