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Bacillus subtilis SepF Binds to the C-Terminus of FtsZ

Bacterial cell division is mediated by a multi-protein machine known as the “divisome”, which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure b...

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Autores principales: Cendrowicz, Ewa, van Kessel, Sebastiaan P., van Bezouwen, Laura S., Kumar, Neeraj, Boekema, Egbert J., Scheffers, Dirk-Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418248/
https://www.ncbi.nlm.nih.gov/pubmed/22912848
http://dx.doi.org/10.1371/journal.pone.0043293
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author Cendrowicz, Ewa
van Kessel, Sebastiaan P.
van Bezouwen, Laura S.
Kumar, Neeraj
Boekema, Egbert J.
Scheffers, Dirk-Jan
author_facet Cendrowicz, Ewa
van Kessel, Sebastiaan P.
van Bezouwen, Laura S.
Kumar, Neeraj
Boekema, Egbert J.
Scheffers, Dirk-Jan
author_sort Cendrowicz, Ewa
collection PubMed
description Bacterial cell division is mediated by a multi-protein machine known as the “divisome”, which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure bacteria divide at the right time and place. Several proteins bind to the Z-ring to mediate its membrane association and persistence throughout the division process. A conserved stretch of amino acids at the C-terminus of FtsZ appears to be involved in many interactions with other proteins. Here, we describe a novel pull-down assay to look for binding partners of the FtsZ C-terminus, using a HaloTag affinity tag fused to the C-terminal 69 amino acids of B. subtilis FtsZ. Using lysates of Escherichia coli overexpressing several B. subtilis cell division proteins as prey we show that the FtsZ C-terminus specifically pulls down SepF, but not EzrA or MinC, and that the interaction depends on a conserved 16 amino acid stretch at the extreme C-terminus. In a reverse pull-down SepF binds to full-length FtsZ but not to a FtsZΔC16 truncate or FtsZ with a mutation of a conserved proline in the C-terminus. We show that the FtsZ C-terminus is required for the formation of tubules from FtsZ polymers by SepF rings. An alanine-scan of the conserved 16 amino acid stretch shows that many mutations affect SepF binding. Combined with the observation that SepF also interacts with the C-terminus of E. coli FtsZ, which is not an in vivo binding partner, we propose that the secondary and tertiary structure of the FtsZ C-terminus, rather than specific amino acids, are recognized by SepF.
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spelling pubmed-34182482012-08-21 Bacillus subtilis SepF Binds to the C-Terminus of FtsZ Cendrowicz, Ewa van Kessel, Sebastiaan P. van Bezouwen, Laura S. Kumar, Neeraj Boekema, Egbert J. Scheffers, Dirk-Jan PLoS One Research Article Bacterial cell division is mediated by a multi-protein machine known as the “divisome”, which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure bacteria divide at the right time and place. Several proteins bind to the Z-ring to mediate its membrane association and persistence throughout the division process. A conserved stretch of amino acids at the C-terminus of FtsZ appears to be involved in many interactions with other proteins. Here, we describe a novel pull-down assay to look for binding partners of the FtsZ C-terminus, using a HaloTag affinity tag fused to the C-terminal 69 amino acids of B. subtilis FtsZ. Using lysates of Escherichia coli overexpressing several B. subtilis cell division proteins as prey we show that the FtsZ C-terminus specifically pulls down SepF, but not EzrA or MinC, and that the interaction depends on a conserved 16 amino acid stretch at the extreme C-terminus. In a reverse pull-down SepF binds to full-length FtsZ but not to a FtsZΔC16 truncate or FtsZ with a mutation of a conserved proline in the C-terminus. We show that the FtsZ C-terminus is required for the formation of tubules from FtsZ polymers by SepF rings. An alanine-scan of the conserved 16 amino acid stretch shows that many mutations affect SepF binding. Combined with the observation that SepF also interacts with the C-terminus of E. coli FtsZ, which is not an in vivo binding partner, we propose that the secondary and tertiary structure of the FtsZ C-terminus, rather than specific amino acids, are recognized by SepF. Public Library of Science 2012-08-13 /pmc/articles/PMC3418248/ /pubmed/22912848 http://dx.doi.org/10.1371/journal.pone.0043293 Text en https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Cendrowicz, Ewa
van Kessel, Sebastiaan P.
van Bezouwen, Laura S.
Kumar, Neeraj
Boekema, Egbert J.
Scheffers, Dirk-Jan
Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title_full Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title_fullStr Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title_full_unstemmed Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title_short Bacillus subtilis SepF Binds to the C-Terminus of FtsZ
title_sort bacillus subtilis sepf binds to the c-terminus of ftsz
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418248/
https://www.ncbi.nlm.nih.gov/pubmed/22912848
http://dx.doi.org/10.1371/journal.pone.0043293
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