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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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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. |
format | Online Article Text |
id | pubmed-3418248 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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