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Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor

Natural competence is the term used to describe the uptake of “naked” extracellular DNA by bacteria; it plays a significant role in horizontal genetic exchange. It is associated with type IV pili, and specialized competence pili mediate DNA uptake. Here, we show that the crystal structure of a compe...

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Autores principales: Salleh, Mohd Zulkifli, Karuppiah, Vijaykumar, Snee, Matthew, Thistlethwaite, Angela, Levy, Colin W., Knight, David, Derrick, Jeremy P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561018/
https://www.ncbi.nlm.nih.gov/pubmed/31186316
http://dx.doi.org/10.1128/mBio.00614-19
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author Salleh, Mohd Zulkifli
Karuppiah, Vijaykumar
Snee, Matthew
Thistlethwaite, Angela
Levy, Colin W.
Knight, David
Derrick, Jeremy P.
author_facet Salleh, Mohd Zulkifli
Karuppiah, Vijaykumar
Snee, Matthew
Thistlethwaite, Angela
Levy, Colin W.
Knight, David
Derrick, Jeremy P.
author_sort Salleh, Mohd Zulkifli
collection PubMed
description Natural competence is the term used to describe the uptake of “naked” extracellular DNA by bacteria; it plays a significant role in horizontal genetic exchange. It is associated with type IV pili, and specialized competence pili mediate DNA uptake. Here, we show that the crystal structure of a competence-associated protein from Thermus thermophilus, ComZ, consists of a type II secretion pseudopilin-like domain, with a large β-solenoid domain inserted into the β-sheet of the pilin-like fold. ComZ binds with high affinity to another competence-associated pilin, PilA2, which lies adjacent to the comZ gene in the genome. The crystal structure of PilA2 revealed a similar type II secretion pseudopilin-like fold, with a small subdomain; docking simulations predicted that PilA2 binds between the pseudopilin-like and β-solenoid domains of ComZ. Electrophoretic shift analysis and DNase protection studies were used to show that ComZ alone and the ComZ/PilA2 complex are able to bind DNA. Protection against reductive dimethylation was used in combination with mass spectrometry and site-directed mutagenesis to identify two lysine residues in ComZ which are involved in DNA binding. They are located between the two domains in ComZ, on the opposite side from the predicted PilA2 binding site. These results suggest a model in which PilA2 assists ComZ in forming the competence pilus tip and DNA binds to the side of the fiber. The results demonstrate how a type IV pilin can be adapted to a specific function by domain insertion and provide the first structural insights into a tip-located competence pilin.
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spelling pubmed-65610182019-06-14 Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor Salleh, Mohd Zulkifli Karuppiah, Vijaykumar Snee, Matthew Thistlethwaite, Angela Levy, Colin W. Knight, David Derrick, Jeremy P. mBio Research Article Natural competence is the term used to describe the uptake of “naked” extracellular DNA by bacteria; it plays a significant role in horizontal genetic exchange. It is associated with type IV pili, and specialized competence pili mediate DNA uptake. Here, we show that the crystal structure of a competence-associated protein from Thermus thermophilus, ComZ, consists of a type II secretion pseudopilin-like domain, with a large β-solenoid domain inserted into the β-sheet of the pilin-like fold. ComZ binds with high affinity to another competence-associated pilin, PilA2, which lies adjacent to the comZ gene in the genome. The crystal structure of PilA2 revealed a similar type II secretion pseudopilin-like fold, with a small subdomain; docking simulations predicted that PilA2 binds between the pseudopilin-like and β-solenoid domains of ComZ. Electrophoretic shift analysis and DNase protection studies were used to show that ComZ alone and the ComZ/PilA2 complex are able to bind DNA. Protection against reductive dimethylation was used in combination with mass spectrometry and site-directed mutagenesis to identify two lysine residues in ComZ which are involved in DNA binding. They are located between the two domains in ComZ, on the opposite side from the predicted PilA2 binding site. These results suggest a model in which PilA2 assists ComZ in forming the competence pilus tip and DNA binds to the side of the fiber. The results demonstrate how a type IV pilin can be adapted to a specific function by domain insertion and provide the first structural insights into a tip-located competence pilin. American Society for Microbiology 2019-06-11 /pmc/articles/PMC6561018/ /pubmed/31186316 http://dx.doi.org/10.1128/mBio.00614-19 Text en Copyright © 2019 Salleh et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Salleh, Mohd Zulkifli
Karuppiah, Vijaykumar
Snee, Matthew
Thistlethwaite, Angela
Levy, Colin W.
Knight, David
Derrick, Jeremy P.
Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title_full Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title_fullStr Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title_full_unstemmed Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title_short Structure and Properties of a Natural Competence-Associated Pilin Suggest a Unique Pilus Tip-Associated DNA Receptor
title_sort structure and properties of a natural competence-associated pilin suggest a unique pilus tip-associated dna receptor
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561018/
https://www.ncbi.nlm.nih.gov/pubmed/31186316
http://dx.doi.org/10.1128/mBio.00614-19
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