The molecular architecture of engulfment during Bacillus subtilis sporulation

The study of bacterial cell biology is limited by difficulties in visualizing cellular structures at high spatial resolution within their native milieu. Here, we visualize Bacillus subtilis sporulation using cryo-electron tomography coupled with cryo-focused ion beam milling, allowing the reconstruc...

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Main Authors: Kanika Khanna, Javier Lopez-Garrido, Ziyi Zhao, Reika Watanabe, Yuan Yuan, Joseph Sugie, Kit Pogliano, Elizabeth Villa
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2019-07-01
Series:eLife
Subjects:
Online Access:https://elifesciences.org/articles/45257
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spelling doaj-56d2c41b0fad498fa79b18682d93915d2021-05-05T17:45:08ZengeLife Sciences Publications LtdeLife2050-084X2019-07-01810.7554/eLife.45257The molecular architecture of engulfment during Bacillus subtilis sporulationKanika Khanna0https://orcid.org/0000-0001-7150-0350Javier Lopez-Garrido1https://orcid.org/0000-0002-8907-502XZiyi Zhao2https://orcid.org/0000-0003-4455-3224Reika Watanabe3https://orcid.org/0000-0002-7427-7702Yuan Yuan4https://orcid.org/0000-0002-9509-9167Joseph Sugie5https://orcid.org/0000-0003-2911-1807Kit Pogliano6https://orcid.org/0000-0002-7868-3345Elizabeth Villa7https://orcid.org/0000-0003-4677-9809Division of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesDivision of Biological Sciences, University of California, San Diego, La Jolla, United StatesThe study of bacterial cell biology is limited by difficulties in visualizing cellular structures at high spatial resolution within their native milieu. Here, we visualize Bacillus subtilis sporulation using cryo-electron tomography coupled with cryo-focused ion beam milling, allowing the reconstruction of native-state cellular sections at molecular resolution. During sporulation, an asymmetrically-positioned septum generates a larger mother cell and a smaller forespore. Subsequently, the mother cell engulfs the forespore. We show that the septal peptidoglycan is not completely degraded at the onset of engulfment. Instead, the septum is uniformly and only slightly thinned as it curves towards the mother cell. Then, the mother cell membrane migrates around the forespore in tiny finger-like projections, whose formation requires the mother cell SpoIIDMP protein complex. We propose that a limited number of SpoIIDMP complexes tether to and degrade the peptidoglycan ahead of the engulfing membrane, generating an irregular membrane front.https://elifesciences.org/articles/45257engulfmentspoIIDMPmembrane migrationpeptidoglycancryo-focused ion beam millingcryo-electron tomography
collection DOAJ
language English
format Article
sources DOAJ
author Kanika Khanna
Javier Lopez-Garrido
Ziyi Zhao
Reika Watanabe
Yuan Yuan
Joseph Sugie
Kit Pogliano
Elizabeth Villa
spellingShingle Kanika Khanna
Javier Lopez-Garrido
Ziyi Zhao
Reika Watanabe
Yuan Yuan
Joseph Sugie
Kit Pogliano
Elizabeth Villa
The molecular architecture of engulfment during Bacillus subtilis sporulation
eLife
engulfment
spoIIDMP
membrane migration
peptidoglycan
cryo-focused ion beam milling
cryo-electron tomography
author_facet Kanika Khanna
Javier Lopez-Garrido
Ziyi Zhao
Reika Watanabe
Yuan Yuan
Joseph Sugie
Kit Pogliano
Elizabeth Villa
author_sort Kanika Khanna
title The molecular architecture of engulfment during Bacillus subtilis sporulation
title_short The molecular architecture of engulfment during Bacillus subtilis sporulation
title_full The molecular architecture of engulfment during Bacillus subtilis sporulation
title_fullStr The molecular architecture of engulfment during Bacillus subtilis sporulation
title_full_unstemmed The molecular architecture of engulfment during Bacillus subtilis sporulation
title_sort molecular architecture of engulfment during bacillus subtilis sporulation
publisher eLife Sciences Publications Ltd
series eLife
issn 2050-084X
publishDate 2019-07-01
description The study of bacterial cell biology is limited by difficulties in visualizing cellular structures at high spatial resolution within their native milieu. Here, we visualize Bacillus subtilis sporulation using cryo-electron tomography coupled with cryo-focused ion beam milling, allowing the reconstruction of native-state cellular sections at molecular resolution. During sporulation, an asymmetrically-positioned septum generates a larger mother cell and a smaller forespore. Subsequently, the mother cell engulfs the forespore. We show that the septal peptidoglycan is not completely degraded at the onset of engulfment. Instead, the septum is uniformly and only slightly thinned as it curves towards the mother cell. Then, the mother cell membrane migrates around the forespore in tiny finger-like projections, whose formation requires the mother cell SpoIIDMP protein complex. We propose that a limited number of SpoIIDMP complexes tether to and degrade the peptidoglycan ahead of the engulfing membrane, generating an irregular membrane front.
topic engulfment
spoIIDMP
membrane migration
peptidoglycan
cryo-focused ion beam milling
cryo-electron tomography
url https://elifesciences.org/articles/45257
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