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

ABSTRACT A gene (comC) essential for natural transformation was identified in Acinetobacter sp. strain BD413. ComC has a typical leader sequence and is similar to different type IV pilus assembly factors. A comC mutant (T308) is not able to bind or take up DNA but exhibits a piliation phenotype indistinguishable from the transformation wild type as revealed by electron microscopy.

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A Type IV Pilus Mediates DNA Binding during Natural Transformation in Streptococcus pneumoniae

PLoS Pathogens ◽

10.1371/journal.ppat.1003473 ◽

2013 ◽

Vol 9 (6) ◽

pp. e1003473 ◽

Cited By ~ 98

Author(s):

Raphaël Laurenceau ◽

Gérard Péhau-Arnaudet ◽

Sonia Baconnais ◽

Joseph Gault ◽

Christian Malosse ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Dna Binding ◽

Natural Transformation ◽

Type Iv Pilus ◽

Type Iv

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Faculty Opinions recommendation of A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae.

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

10.3410/f.718046837.793481030 ◽

2013 ◽

Author(s):

David Stephens ◽

Scott Chancey

Keyword(s):

Streptococcus Pneumoniae ◽

Dna Binding ◽

Natural Transformation ◽

Type Iv Pilus ◽

Type Iv

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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

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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

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DNA Binding: a Novel Function of Pseudomonas aeruginosa Type IV Pili

Journal of Bacteriology ◽

10.1128/jb.187.4.1455-1464.2005 ◽

2005 ◽

Vol 187 (4) ◽

pp. 1455-1464 ◽

Cited By ~ 96

Author(s):

Erin J. van Schaik ◽

Carmen L. Giltner ◽

Gerald F. Audette ◽

David W. Keizer ◽

Daisy L. Bautista ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Dna Binding ◽

Biofilm Formation ◽

Quaternary Structure ◽

Natural Transformation ◽

Ex Vivo ◽

Specific Binding ◽

Type Iv Pili ◽

Type Iv ◽

Bacteriophage Infection

ABSTRACT The opportunistic pathogen Pseudomonas aeruginosa produces multifunctional, polar, filamentous appendages termed type IV pili. Type IV pili are involved in colonization during infection, twitching motility, biofilm formation, bacteriophage infection, and natural transformation. Electrostatic surface analysis of modeled pilus fibers generated from P. aeruginosa strain PAK, K122-4, and KB-7 pilin monomers suggested that a solvent-exposed band of positive charge may be a common feature of all type IV pili. Several functions of type IV pili, including natural transformation and biofilm formation, involve DNA. We investigated the ability of P. aeruginosa type IV pili to bind DNA. Purified PAK, K122-4, and KB-7 pili were observed to bind both bacterial plasmid and salmon sperm DNA in a concentration-dependent and saturable manner. PAK pili had the highest affinity for DNA, followed by K122-4 and KB-7 pili. DNA binding involved backbone interactions and preferential binding to pyrimidine residues even though there was no evidence of sequence-specific binding. Pilus-mediated DNA binding was a function of the intact pilus and thus required elements present in the quaternary structure. However, binding also involved the pilus tip as tip-specific, but not base-specific, antibodies inhibited DNA binding. The conservation of a Thr residue in all type IV pilin monomers examined to date, along with the electrostatic data, implies that DNA binding is a conserved function of type IV pili. Pilus-mediated DNA binding could be important for biofilm formation both in vivo during an infection and ex vivo on abiotic surfaces.

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Discovery of a new Neisseria gonorrhoeae Type IV pilus assembly factor, TfpC

10.1101/2020.09.26.314724 ◽

2020 ◽

Author(s):

Linda I. Hu ◽

Shaohui Yin ◽

Egon A. Ozer ◽

Lee Sewell ◽

Saima Rehman ◽

...

Keyword(s):

Cell Surface ◽

Neisseria Gonorrhoeae ◽

Sexually Transmitted Infection ◽

Bacterial Cell ◽

Bacterial Species ◽

Type Iv Pili ◽

Type Iv Pilus ◽

Transmitted Infection ◽

Sexually Transmitted ◽

Type Iv

AbstractNeisseria gonorrhoeae rely on Type IV pili (T4p) to promote colonization of their human host and to cause the sexually transmitted infection, gonorrhea. This organelle cycles through a process of extension and retraction back into the bacterial cell. Through a genetic screen, we identified the NGO0783 locus of N. gonorrhoeae strain FA1090 as containing a gene encoding a protein required to stabilize the Type IV pilus in its extended, non-retracted conformation. We have named the gene tfpC and the protein TfpC. Deletion of tfpC produces a nonpiliated colony morphology and immuno-transmission electron microscopy confirms that the pili are lost in the ΔtfpC mutant, although there is some pilin detected near the bacterial cell surface. A copy of the tfpC gene expressed from a lac promoter restores pilus expression and related phenotypes. A ΔtfpC mutant shows reduced levels of pilin protein, but complementation with a tfpC gene restored pilin to normal levels. Bioinformatic searches show there are orthologues in numerous bacteria species but not all Type IV pilin expressing bacteria contain orthologous genes. Co-evolution and NMR analysis indicates that TfpC contains an N-terminal transmembrane helix, a substantial extended/unstructured region and a highly charge C-terminal coiled-coil domain.ImportanceMost bacterial species express one or more extracellular organelles called pili/fimbriae that are required for many properties of each bacterial cell. The Neisseria gonorrhoeae Type IV pilus is a major virulence and colonization factor for the sexually transmitted infection, gonorrhea. We have discovered a new protein of Neisseria gonorrhoeae called TfpC that is required to maintain the Type IV pili on the bacterial cell surface. There are similar proteins found in the other members of the Neisseria genus and many other bacterial species important for human health.

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Discovery of a New Neisseria gonorrhoeae Type IV Pilus Assembly Factor, TfpC

mBio ◽

10.1128/mbio.02528-20 ◽

2020 ◽

Vol 11 (5) ◽

Author(s):

Linda I. Hu ◽

Shaohui Yin ◽

Egon A. Ozer ◽

Lee Sewell ◽

Saima Rehman ◽

...

Keyword(s):

Cell Surface ◽

Neisseria Gonorrhoeae ◽

Bacterial Cell ◽

Bacterial Species ◽

Type Iv Pili ◽

Type Iv Pilus ◽

Transmitted Infection ◽

Content Type ◽

Sexually Transmitted ◽

Type Iv

ABSTRACT Neisseria gonorrhoeae relies on type IV pili (T4p) to promote colonization of their human host and to cause the sexually transmitted infection gonorrhea. This organelle cycles through a process of extension and retraction back into the bacterial cell. Through a genetic screen, we identified the NGO0783 locus of N. gonorrhoeae strain FA1090 as containing a gene encoding a protein required to stabilize the type IV pilus in its extended, nonretracted conformation. We have named the gene tfpC and the protein TfpC. Deletion of tfpC produces a nonpiliated colony morphology, and immuno-transmission electron microscopy confirms that the pili are lost in the ΔtfpC mutant, although there is some pilin detected near the bacterial cell surface. A copy of the tfpC gene expressed from a lac promoter restores pilus expression and related phenotypes. A ΔtfpC mutant shows reduced levels of pilin protein, but complementation with a tfpC gene restored pilin to normal levels. Bioinformatic searches show that there are orthologues in numerous bacterial species, but not all type IV pilin-expressing bacteria contain orthologous genes. Coevolution and nuclear magnetic resonance (NMR) analysis indicates that TfpC contains an N-terminal transmembrane helix, a substantial extended/unstructured region, and a highly charged C-terminal coiled-coil domain. IMPORTANCE Most bacterial species express one or more extracellular organelles called pili/fimbriae that are required for many properties of each bacterial cell. The Neisseria gonorrhoeae type IV pilus is a major virulence and colonization factor for the sexually transmitted infection gonorrhea. We have discovered a new protein of Neisseria gonorrhoeae called TfpC that is required to maintain type IV pili on the bacterial cell surface. There are similar proteins found in other members of the Neisseria genus and many other bacterial species important for human health.

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Attenuation of the Type IV Pilus Retraction Motor Influences Neisseria gonorrhoeae Social and Infection Behavior

mBio ◽

10.1128/mbio.01994-16 ◽

2016 ◽

Vol 7 (6) ◽

Cited By ~ 10

Author(s):

Alyson M. Hockenberry ◽

Danielle M. Hutchens ◽

Al Agellon ◽

Magdalene So

Keyword(s):

Epidermal Growth Factor Receptor ◽

Growth Factor ◽

Social Behavior ◽

Epidermal Growth Factor ◽

Neisseria Gonorrhoeae ◽

Enzymatic Activity ◽

Growth Factor Receptor ◽

Type Iv Pilus ◽

Type Iv ◽

Epidermal Growth

ABSTRACT Retraction of the type IV pilus (Tfp) mediates DNA uptake, motility, and social and infection behavior in a wide variety of prokaryotes. To date, investigations into Tfp retraction-dependent activities have used a mutant deleted of PilT, the ATPase motor protein that causes the pilus fiber to retract. Δ pilT cells are nontransformable, nonmotile, and cannot aggregate into microcolonies. We tested the hypothesis that these retraction-dependent activities are sensitive to the strength of PilT enzymatic activity by using the pathogen Neisseria gonorrhoeae as a model. We constructed an N. gonorrhoeae mutant with an amino acid substitution in the PilT Walker B box (a substitution of cysteine for leucine at position 201, encoded by pilT L201C ). Purified PilT L201C forms a native hexamer, but mutant hexamers hydrolyze ATP at half the maximal rate. N. gonorrhoeae pilT L201C cells produce Tfp fibers, crawl at the same speed as the wild-type (wt) parent, and are equally transformable. However, the social behavior of pilT L201C cells is intermediate between the behaviors of wt and Δ pilT cells. The infection behavior of pilT L201C is also defective, due to its failure to activate the epidermal growth factor receptor (EGFR)-heparin-binding EGF-like growth factor (HB-EGF) pathway. Our study indicates that pilus retraction, per se , is not sufficient for N. gonorrhoeae microcolony formation or infectivity; rather, these activities are sensitive to the strength of PilT enzymatic activity. We discuss the implications of these findings for Neisseria pathogenesis in the context of mechanobiology. IMPORTANCE Type IV pili are fibers expressed on the surface of many bacteria. Neisseria gonorrhoeae cells crawl, take up DNA, and communicate with each other and with human cells by retracting these fibers. Here, we show that an N. gonorrhoeae mutant expressing an enzymatically weakened type IV pilus retraction motor still crawls and takes up DNA normally. However, mutant cells exhibit abnormal social behavior, and they are less infective because they fail to activate the epidermal growth factor receptor. Our study shows that N. gonorrhoeae social and infection behaviors are sensitive to the strength of the retraction motor enzyme.

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