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Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein

During Bacillus subtilis sporulation, segregating sister chromosomes are anchored to cell poles and the chromosome is remodeled into an elongated structure called the axial filament. Data indicate that a developmentally regulated protein called RacA is involved in these functions. To gain insight in...

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Autores principales: Schumacher, Maria A., Lee, Jeehyun, Zeng, Wenjie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4914108/
https://www.ncbi.nlm.nih.gov/pubmed/27085804
http://dx.doi.org/10.1093/nar/gkw248
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author Schumacher, Maria A.
Lee, Jeehyun
Zeng, Wenjie
author_facet Schumacher, Maria A.
Lee, Jeehyun
Zeng, Wenjie
author_sort Schumacher, Maria A.
collection PubMed
description During Bacillus subtilis sporulation, segregating sister chromosomes are anchored to cell poles and the chromosome is remodeled into an elongated structure called the axial filament. Data indicate that a developmentally regulated protein called RacA is involved in these functions. To gain insight into how RacA performs these diverse processes we performed a battery of structural and biochemical analyses. These studies show that RacA contains an N-terminal winged-helix-turn-helix module connected by a disordered region to a predicted coiled-coil domain. Structures capture RacA binding the DNA using distinct protein–protein interfaces and employing adjustable DNA docking modes. This unique DNA binding mechanism indicates how RacA can both specifically recognize its GC-rich centromere and also non-specifically bind the DNA. Adjacent RacA molecules within the protein–DNA structure interact leading to DNA compaction, suggesting a mechanism for axial filament formation. We also show that the RacA C-domain coiled coil directly contacts the coiled coil region of the polar protein DivIVA, which anchors RacA and hence the chromosome to the pole. Thus, our combined data reveal unique DNA binding properties by RacA and provide insight into the DNA remodeling and polar anchorage functions of the protein.
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spelling pubmed-49141082016-06-22 Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein Schumacher, Maria A. Lee, Jeehyun Zeng, Wenjie Nucleic Acids Res Structural Biology During Bacillus subtilis sporulation, segregating sister chromosomes are anchored to cell poles and the chromosome is remodeled into an elongated structure called the axial filament. Data indicate that a developmentally regulated protein called RacA is involved in these functions. To gain insight into how RacA performs these diverse processes we performed a battery of structural and biochemical analyses. These studies show that RacA contains an N-terminal winged-helix-turn-helix module connected by a disordered region to a predicted coiled-coil domain. Structures capture RacA binding the DNA using distinct protein–protein interfaces and employing adjustable DNA docking modes. This unique DNA binding mechanism indicates how RacA can both specifically recognize its GC-rich centromere and also non-specifically bind the DNA. Adjacent RacA molecules within the protein–DNA structure interact leading to DNA compaction, suggesting a mechanism for axial filament formation. We also show that the RacA C-domain coiled coil directly contacts the coiled coil region of the polar protein DivIVA, which anchors RacA and hence the chromosome to the pole. Thus, our combined data reveal unique DNA binding properties by RacA and provide insight into the DNA remodeling and polar anchorage functions of the protein. Oxford University Press 2016-06-20 2016-04-16 /pmc/articles/PMC4914108/ /pubmed/27085804 http://dx.doi.org/10.1093/nar/gkw248 Text en © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Structural Biology
Schumacher, Maria A.
Lee, Jeehyun
Zeng, Wenjie
Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title_full Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title_fullStr Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title_full_unstemmed Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title_short Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein
title_sort molecular insights into dna binding and anchoring by the bacillus subtilis sporulation kinetochore-like raca protein
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4914108/
https://www.ncbi.nlm.nih.gov/pubmed/27085804
http://dx.doi.org/10.1093/nar/gkw248
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