Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>

ABSTRACT The assembly of the cell division machinery at midcell is a critical step of cytokinesis. Many rod-shaped bacteria position septa using nucleoid occlusion, which prevents division over the chromosome, and the Min system, which prevents division near the poles. Here we examined the in vivo a...

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Autores principales:	Prahathees Eswaramoorthy, Marcella L. Erb, James A. Gregory, Jared Silverman, Kit Pogliano, Joe Pogliano, Kumaran S. Ramamurthi
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2011
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ec3f844659f44d30b8561ecc32ef79a6
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spelling	oai:doaj.org-article:ec3f844659f44d30b8561ecc32ef79a62021-11-15T15:38:48ZCellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>10.1128/mBio.00257-112150-7511https://doaj.org/article/ec3f844659f44d30b8561ecc32ef79a62011-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00257-11https://doaj.org/toc/2150-7511ABSTRACT The assembly of the cell division machinery at midcell is a critical step of cytokinesis. Many rod-shaped bacteria position septa using nucleoid occlusion, which prevents division over the chromosome, and the Min system, which prevents division near the poles. Here we examined the in vivo assembly of the Bacillus subtilis MinCD targeting proteins DivIVA, a peripheral membrane protein that preferentially localizes to negatively curved membranes and resembles eukaryotic tropomyosins, and MinJ, which recruits MinCD to DivIVA. We used structured illumination microscopy to demonstrate that both DivIVA and MinJ localize as double rings that flank the septum and first appear early in septal biosynthesis. The subsequent recruitment of MinCD to these double rings would separate the Min proteins from their target, FtsZ, spatially regulating Min activity and allowing continued cell division. Curvature-based localization would also provide temporal regulation, since DivIVA and the Min proteins would localize to midcell after the onset of division. We use time-lapse microscopy and fluorescence recovery after photobleaching to demonstrate that DivIVA rings are highly stable and are constructed from newly synthesized DivIVA molecules. After cell division, DivIVA rings appear to collapse into patches at the rounded cell poles of separated cells, with little or no incorporation of newly synthesized subunits. Thus, changes in cell architecture mediate both the initial recruitment of DivIVA to sites of cell division and the subsequent collapse of these rings into patches (or rings of smaller diameter), while curvature-based localization of DivIVA spatially and temporally regulates Min activity. IMPORTANCE The Min systems of Escherichia coli and Bacillus subtilis both inhibit FtsZ assembly, but one key difference between these two species is that whereas the E. coli Min proteins localize to the poles, the B. subtilis proteins localize to nascent division sites by interaction with DivIVA and MinJ. It is unclear how MinC activity at midcell is regulated to prevent it from interfering with FtsZ engaged in medial cell division. We used superresolution microscopy to demonstrate that DivIVA and MinJ, which localize MinCD, assemble double rings that flank active division sites and septa. This curvature-based localization mechanism holds MinCD away from the FtsZ ring at midcell, and we propose that this spatial organization is the primary mechanism by which MinC activity is regulated to allow division at midcell. Curvature-based localization also conveys temporal regulation, since it ensures that MinC localizes after the onset of division.Prahathees EswaramoorthyMarcella L. ErbJames A. GregoryJared SilvermanKit PoglianoJoe PoglianoKumaran S. RamamurthiAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 6 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Prahathees Eswaramoorthy Marcella L. Erb James A. Gregory Jared Silverman Kit Pogliano Joe Pogliano Kumaran S. Ramamurthi Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
description	ABSTRACT The assembly of the cell division machinery at midcell is a critical step of cytokinesis. Many rod-shaped bacteria position septa using nucleoid occlusion, which prevents division over the chromosome, and the Min system, which prevents division near the poles. Here we examined the in vivo assembly of the Bacillus subtilis MinCD targeting proteins DivIVA, a peripheral membrane protein that preferentially localizes to negatively curved membranes and resembles eukaryotic tropomyosins, and MinJ, which recruits MinCD to DivIVA. We used structured illumination microscopy to demonstrate that both DivIVA and MinJ localize as double rings that flank the septum and first appear early in septal biosynthesis. The subsequent recruitment of MinCD to these double rings would separate the Min proteins from their target, FtsZ, spatially regulating Min activity and allowing continued cell division. Curvature-based localization would also provide temporal regulation, since DivIVA and the Min proteins would localize to midcell after the onset of division. We use time-lapse microscopy and fluorescence recovery after photobleaching to demonstrate that DivIVA rings are highly stable and are constructed from newly synthesized DivIVA molecules. After cell division, DivIVA rings appear to collapse into patches at the rounded cell poles of separated cells, with little or no incorporation of newly synthesized subunits. Thus, changes in cell architecture mediate both the initial recruitment of DivIVA to sites of cell division and the subsequent collapse of these rings into patches (or rings of smaller diameter), while curvature-based localization of DivIVA spatially and temporally regulates Min activity. IMPORTANCE The Min systems of Escherichia coli and Bacillus subtilis both inhibit FtsZ assembly, but one key difference between these two species is that whereas the E. coli Min proteins localize to the poles, the B. subtilis proteins localize to nascent division sites by interaction with DivIVA and MinJ. It is unclear how MinC activity at midcell is regulated to prevent it from interfering with FtsZ engaged in medial cell division. We used superresolution microscopy to demonstrate that DivIVA and MinJ, which localize MinCD, assemble double rings that flank active division sites and septa. This curvature-based localization mechanism holds MinCD away from the FtsZ ring at midcell, and we propose that this spatial organization is the primary mechanism by which MinC activity is regulated to allow division at midcell. Curvature-based localization also conveys temporal regulation, since it ensures that MinC localizes after the onset of division.
format	article
author	Prahathees Eswaramoorthy Marcella L. Erb James A. Gregory Jared Silverman Kit Pogliano Joe Pogliano Kumaran S. Ramamurthi
author_facet	Prahathees Eswaramoorthy Marcella L. Erb James A. Gregory Jared Silverman Kit Pogliano Joe Pogliano Kumaran S. Ramamurthi
author_sort	Prahathees Eswaramoorthy
title	Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_short	Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_full	Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_fullStr	Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_full_unstemmed	Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>
title_sort	cellular architecture mediates diviva ultrastructure and regulates min activity in <named-content content-type="genus-species">bacillus subtilis</named-content>
publisher	American Society for Microbiology
publishDate	2011
url	https://doaj.org/article/ec3f844659f44d30b8561ecc32ef79a6
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Cellular Architecture Mediates DivIVA Ultrastructure and Regulates Min Activity in <named-content content-type="genus-species">Bacillus subtilis</named-content>

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