Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf
Shewanella vesiculosa M7 T is a cold-adapted Antarctic bacterium that has a great capacity to secrete membrane vesicles (MVs), making it a potentially excellent model for studying the vesiculation process. S. vesiculosa M7 T undergoes a blebbing mechanism to produce different types of MVs, including...
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ftfrontimediafig:oai:figshare.com:article/16756846 2023-05-15T14:02:18+02:00 Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf Nicolás Baeza Lidia Delgado Jaume Comas Elena Mercade 2021-10-07T04:44:26Z https://doi.org/10.3389/fmicb.2021.713669.s001 https://figshare.com/articles/presentation/Presentation_1_Phage-Mediated_Explosive_Cell_Lysis_Induces_the_Formation_of_a_Different_Type_of_O-IMV_in_Shewanella_vesiculosa_M7T_pdf/16756846 unknown doi:10.3389/fmicb.2021.713669.s001 https://figshare.com/articles/presentation/Presentation_1_Phage-Mediated_Explosive_Cell_Lysis_Induces_the_Formation_of_a_Different_Type_of_O-IMV_in_Shewanella_vesiculosa_M7T_pdf/16756846 CC BY 4.0 CC-BY Microbiology Microbial Genetics Microbial Ecology Mycology membrane vesicles gram-negative bacteria explosive cell lysis OMV O-IMV DNA flow cytometry electron microscopy Text Presentation 2021 ftfrontimediafig https://doi.org/10.3389/fmicb.2021.713669.s001 2021-10-13T23:02:47Z Shewanella vesiculosa M7 T is a cold-adapted Antarctic bacterium that has a great capacity to secrete membrane vesicles (MVs), making it a potentially excellent model for studying the vesiculation process. S. vesiculosa M7 T undergoes a blebbing mechanism to produce different types of MVs, including outer membrane vesicles and outer-inner membrane vesicles (O-IMVs). More recently, other mechanisms have been considered that could lead to the formation of O-IMVs derived from prophage-mediated explosive cell lysis in other bacteria, but it is not clear if they are of the same type. The bacterial growth phase could also have a great impact on the type of MVs, although there are few studies on the subject. In this study, we used high-resolution flow cytometry, transmission electron microscopy, and cryo-electron microscopy (Cryo-EM) analysis to determine the amount and types of MVs S. vesiculosa M7 T secreted during different growth phases. We show that MV secretion increases during the transition from the late exponential to the stationary phase. Moreover, prophage-mediated explosive cell lysis is activated in S. vesiculosa M7 T , increasing the heterogeneity of both single- and double-layer MVs. The sequenced DNA fragments from the MVs covered the entire genome, confirming this explosive cell lysis mechanism. A different structure and biogenesis mechanisms for the explosive cell lysis-derived double-layered MVs was observed, and we propose to name them explosive O-IMVs, distinguishing them from the blebbing O-IMVs; their separation is a first step to elucidate their different functions. In our study, we used for the first time sorting by flow cytometry and Cryo-EM analyses to isolate bacterial MVs based on their nucleic acid content. Further improvements and implementation of bacterial MV separation techniques is essential to develop more in-depth knowledge of MVs. Conference Object Antarc* Antarctic Frontiers: Figshare Antarctic |
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Frontiers: Figshare |
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ftfrontimediafig |
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unknown |
topic |
Microbiology Microbial Genetics Microbial Ecology Mycology membrane vesicles gram-negative bacteria explosive cell lysis OMV O-IMV DNA flow cytometry electron microscopy |
spellingShingle |
Microbiology Microbial Genetics Microbial Ecology Mycology membrane vesicles gram-negative bacteria explosive cell lysis OMV O-IMV DNA flow cytometry electron microscopy Nicolás Baeza Lidia Delgado Jaume Comas Elena Mercade Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
topic_facet |
Microbiology Microbial Genetics Microbial Ecology Mycology membrane vesicles gram-negative bacteria explosive cell lysis OMV O-IMV DNA flow cytometry electron microscopy |
description |
Shewanella vesiculosa M7 T is a cold-adapted Antarctic bacterium that has a great capacity to secrete membrane vesicles (MVs), making it a potentially excellent model for studying the vesiculation process. S. vesiculosa M7 T undergoes a blebbing mechanism to produce different types of MVs, including outer membrane vesicles and outer-inner membrane vesicles (O-IMVs). More recently, other mechanisms have been considered that could lead to the formation of O-IMVs derived from prophage-mediated explosive cell lysis in other bacteria, but it is not clear if they are of the same type. The bacterial growth phase could also have a great impact on the type of MVs, although there are few studies on the subject. In this study, we used high-resolution flow cytometry, transmission electron microscopy, and cryo-electron microscopy (Cryo-EM) analysis to determine the amount and types of MVs S. vesiculosa M7 T secreted during different growth phases. We show that MV secretion increases during the transition from the late exponential to the stationary phase. Moreover, prophage-mediated explosive cell lysis is activated in S. vesiculosa M7 T , increasing the heterogeneity of both single- and double-layer MVs. The sequenced DNA fragments from the MVs covered the entire genome, confirming this explosive cell lysis mechanism. A different structure and biogenesis mechanisms for the explosive cell lysis-derived double-layered MVs was observed, and we propose to name them explosive O-IMVs, distinguishing them from the blebbing O-IMVs; their separation is a first step to elucidate their different functions. In our study, we used for the first time sorting by flow cytometry and Cryo-EM analyses to isolate bacterial MVs based on their nucleic acid content. Further improvements and implementation of bacterial MV separation techniques is essential to develop more in-depth knowledge of MVs. |
format |
Conference Object |
author |
Nicolás Baeza Lidia Delgado Jaume Comas Elena Mercade |
author_facet |
Nicolás Baeza Lidia Delgado Jaume Comas Elena Mercade |
author_sort |
Nicolás Baeza |
title |
Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
title_short |
Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
title_full |
Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
title_fullStr |
Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
title_full_unstemmed |
Presentation_1_Phage-Mediated Explosive Cell Lysis Induces the Formation of a Different Type of O-IMV in Shewanella vesiculosa M7T.pdf |
title_sort |
presentation_1_phage-mediated explosive cell lysis induces the formation of a different type of o-imv in shewanella vesiculosa m7t.pdf |
publishDate |
2021 |
url |
https://doi.org/10.3389/fmicb.2021.713669.s001 https://figshare.com/articles/presentation/Presentation_1_Phage-Mediated_Explosive_Cell_Lysis_Induces_the_Formation_of_a_Different_Type_of_O-IMV_in_Shewanella_vesiculosa_M7T_pdf/16756846 |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
doi:10.3389/fmicb.2021.713669.s001 https://figshare.com/articles/presentation/Presentation_1_Phage-Mediated_Explosive_Cell_Lysis_Induces_the_Formation_of_a_Different_Type_of_O-IMV_in_Shewanella_vesiculosa_M7T_pdf/16756846 |
op_rights |
CC BY 4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fmicb.2021.713669.s001 |
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1766272522319298560 |