<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>

ABSTRACT Outer membrane vesicles (OMVs) are universally produced by prokaryotes and play important roles in symbiotic and pathogenic interactions. They often contain DNA, but a mechanism for its incorporation is lacking. Here, we show that Dinoroseobacter shibae, a dinoflagellate symbiont, constitut...

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Autores principales:	Hui Wang, Nicole Beier, Christian Boedeker, Helena Sztajer, Petra Henke, Meina Neumann-Schaal, Johannes Mansky, Manfred Rohde, Jörg Overmann, Jörn Petersen, Frank Klawonn, Martin Kucklick, Susanne Engelmann, Jürgen Tomasch, Irene Wagner-Döbler
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	DNA repair OMV vesicles replication termination circular chromosomes Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b502b34a4bca4dfe8c990c9d2e511736
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spelling	oai:doaj.org-article:b502b34a4bca4dfe8c990c9d2e5117362021-12-02T19:36:37Z<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>10.1128/mSystems.00693-202379-5077https://doaj.org/article/b502b34a4bca4dfe8c990c9d2e5117362021-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00693-20https://doaj.org/toc/2379-5077ABSTRACT Outer membrane vesicles (OMVs) are universally produced by prokaryotes and play important roles in symbiotic and pathogenic interactions. They often contain DNA, but a mechanism for its incorporation is lacking. Here, we show that Dinoroseobacter shibae, a dinoflagellate symbiont, constitutively secretes OMVs containing DNA. Time-lapse microscopy captured instances of multiple OMV production at the septum during cell division. DNA from the vesicle lumen was up to 22-fold enriched for the region around the terminus of replication (ter). The peak of coverage was located at dif, a conserved 28-bp palindromic sequence required for binding of the site-specific tyrosine recombinases XerC/XerD. These enzymes are activated at the last stage of cell division immediately prior to septum formation when they are bound by the divisome protein FtsK. We suggest that overreplicated regions around the terminus have been repaired by the FtsK-dif-XerC/XerD molecular machinery. The vesicle proteome was clearly dominated by outer membrane and periplasmic proteins. Some of the most abundant vesicle membrane proteins were predicted to be required for direct interaction with peptidoglycan during cell division (LysM, Tol-Pal, Spol, lytic murein transglycosylase). OMVs were 15-fold enriched for the saturated fatty acid 16:00. We hypothesize that constitutive OMV secretion in D. shibae is coupled to cell division. The footprint of the FtsK-dif-XerC/XerD molecular machinery suggests a novel potentially highly conserved route for incorporation of DNA into OMVs. Clearing the division site from small DNA fragments might be an important function of vesicles produced during exponential growth under optimal conditions. IMPORTANCE Gram-negative bacteria continually form vesicles from their outer membrane (outer membrane vesicles [OMVs]) during normal growth. OMVs frequently contain DNA, and it is unclear how DNA can be shuffled from the cytoplasm to the OMVs. We studied OMV cargo in Dinoroseobacter shibae, a symbiont of dinoflagellates, using microscopy and a multi-omics approach. We found that vesicles formed during undisturbed exponential growth contain DNA which is enriched for genes around the replication terminus, specifically, the binding site for an enzyme complex that is activated at the last stage of cell division. We suggest that the enriched genes are the result of overreplication which is repaired by their excision and excretion via membrane vesicles to clear the divisome from waste DNA.Hui WangNicole BeierChristian BoedekerHelena SztajerPetra HenkeMeina Neumann-SchaalJohannes ManskyManfred RohdeJörg OvermannJörn PetersenFrank KlawonnMartin KucklickSusanne EngelmannJürgen TomaschIrene Wagner-DöblerAmerican Society for MicrobiologyarticleDNA repairOMVvesiclesreplication terminationcircular chromosomesMicrobiologyQR1-502ENmSystems, Vol 6, Iss 1 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	DNA repair OMV vesicles replication termination circular chromosomes Microbiology QR1-502
spellingShingle	DNA repair OMV vesicles replication termination circular chromosomes Microbiology QR1-502 Hui Wang Nicole Beier Christian Boedeker Helena Sztajer Petra Henke Meina Neumann-Schaal Johannes Mansky Manfred Rohde Jörg Overmann Jörn Petersen Frank Klawonn Martin Kucklick Susanne Engelmann Jürgen Tomasch Irene Wagner-Döbler <named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
description	ABSTRACT Outer membrane vesicles (OMVs) are universally produced by prokaryotes and play important roles in symbiotic and pathogenic interactions. They often contain DNA, but a mechanism for its incorporation is lacking. Here, we show that Dinoroseobacter shibae, a dinoflagellate symbiont, constitutively secretes OMVs containing DNA. Time-lapse microscopy captured instances of multiple OMV production at the septum during cell division. DNA from the vesicle lumen was up to 22-fold enriched for the region around the terminus of replication (ter). The peak of coverage was located at dif, a conserved 28-bp palindromic sequence required for binding of the site-specific tyrosine recombinases XerC/XerD. These enzymes are activated at the last stage of cell division immediately prior to septum formation when they are bound by the divisome protein FtsK. We suggest that overreplicated regions around the terminus have been repaired by the FtsK-dif-XerC/XerD molecular machinery. The vesicle proteome was clearly dominated by outer membrane and periplasmic proteins. Some of the most abundant vesicle membrane proteins were predicted to be required for direct interaction with peptidoglycan during cell division (LysM, Tol-Pal, Spol, lytic murein transglycosylase). OMVs were 15-fold enriched for the saturated fatty acid 16:00. We hypothesize that constitutive OMV secretion in D. shibae is coupled to cell division. The footprint of the FtsK-dif-XerC/XerD molecular machinery suggests a novel potentially highly conserved route for incorporation of DNA into OMVs. Clearing the division site from small DNA fragments might be an important function of vesicles produced during exponential growth under optimal conditions. IMPORTANCE Gram-negative bacteria continually form vesicles from their outer membrane (outer membrane vesicles [OMVs]) during normal growth. OMVs frequently contain DNA, and it is unclear how DNA can be shuffled from the cytoplasm to the OMVs. We studied OMV cargo in Dinoroseobacter shibae, a symbiont of dinoflagellates, using microscopy and a multi-omics approach. We found that vesicles formed during undisturbed exponential growth contain DNA which is enriched for genes around the replication terminus, specifically, the binding site for an enzyme complex that is activated at the last stage of cell division. We suggest that the enriched genes are the result of overreplication which is repaired by their excision and excretion via membrane vesicles to clear the divisome from waste DNA.
format	article
author	Hui Wang Nicole Beier Christian Boedeker Helena Sztajer Petra Henke Meina Neumann-Schaal Johannes Mansky Manfred Rohde Jörg Overmann Jörn Petersen Frank Klawonn Martin Kucklick Susanne Engelmann Jürgen Tomasch Irene Wagner-Döbler
author_facet	Hui Wang Nicole Beier Christian Boedeker Helena Sztajer Petra Henke Meina Neumann-Schaal Johannes Mansky Manfred Rohde Jörg Overmann Jörn Petersen Frank Klawonn Martin Kucklick Susanne Engelmann Jürgen Tomasch Irene Wagner-Döbler
author_sort	Hui Wang
title	<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
title_short	<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
title_full	<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
title_fullStr	<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
title_full_unstemmed	<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>
title_sort	<named-content content-type="genus-species">dinoroseobacter shibae</named-content> outer membrane vesicles are enriched for the chromosome dimer resolution site <italic toggle="yes">dif</italic>
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/b502b34a4bca4dfe8c990c9d2e511736
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<named-content content-type="genus-species">Dinoroseobacter shibae</named-content> Outer Membrane Vesicles Are Enriched for the Chromosome Dimer Resolution Site <italic toggle="yes">dif</italic>

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