Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells

ABSTRACT Type IV secretion systems (T4SS) transfer DNA and/or proteins into recipient cells. Here we performed immunofluorescence deconvolution microscopy to localize the assembled T4SS by detection of its native components VirB1, VirB2, VirB4, VirB5, VirB7, VirB8, VirB9, VirB10, and VirB11 in the C...

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Autores principales:	Julieta Aguilar, Todd A. Cameron, John Zupan, Patricia Zambryski
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2011
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/1d25c74eda8f4a6e873bc9aa4895eb5e
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spelling	oai:doaj.org-article:1d25c74eda8f4a6e873bc9aa4895eb5e2021-11-15T15:38:48ZMembrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells10.1128/mBio.00218-112150-7511https://doaj.org/article/1d25c74eda8f4a6e873bc9aa4895eb5e2011-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00218-11https://doaj.org/toc/2150-7511ABSTRACT Type IV secretion systems (T4SS) transfer DNA and/or proteins into recipient cells. Here we performed immunofluorescence deconvolution microscopy to localize the assembled T4SS by detection of its native components VirB1, VirB2, VirB4, VirB5, VirB7, VirB8, VirB9, VirB10, and VirB11 in the C58 nopaline strain of Agrobacterium tumefaciens, following induction of virulence (vir) gene expression. These different proteins represent T4SS components spanning the inner membrane, periplasm, or outer membrane. Native VirB2, VirB5, VirB7, and VirB8 were also localized in the A. tumefaciens octopine strain A348. Quantitative analyses of the localization of all the above Vir proteins in nopaline and octopine strains revealed multiple foci in single optical sections in over 80% and 70% of the bacterial cells, respectively. Green fluorescent protein (GFP)-VirB8 expression following vir induction was used to monitor bacterial binding to live host plant cells; bacteria bind predominantly along their lengths, with few bacteria binding via their poles or subpoles. vir-induced attachment-defective bacteria or bacteria without the Ti plasmid do not bind to plant cells. These data support a model where multiple vir-T4SS around the perimeter of the bacterium maximize effective contact with the host to facilitate efficient transfer of DNA and protein substrates. IMPORTANCE Transfer of DNA and/or proteins to host cells through multiprotein type IV secretion system (T4SS) complexes that span the bacterial cell envelope is critical to bacterial pathogenesis. Early reports suggested that T4SS components localized at the cell poles. Now, higher-resolution deconvolution fluorescence microscopy reveals that all structural components of the Agrobacterium tumefaciens vir-T4SS, as well as its transported protein substrates, localize to multiple foci around the cell perimeter. These results lead to a new model of A. tumefaciens attachment to a plant cell, where A. tumefaciens takes advantage of the multiple vir-T4SS along its length to make intimate lateral contact with plant cells and thereby effectively transfer DNA and/or proteins through the vir-T4SS. The T4SS of A. tumefaciens is among the best-studied T4SS, and the majority of its components are highly conserved in different pathogenic bacterial species. Thus, the results presented can be applied to a broad range of pathogens that utilize T4SS.Julieta AguilarTodd A. CameronJohn ZupanPatricia ZambryskiAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 2, Iss 6 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Julieta Aguilar Todd A. Cameron John Zupan Patricia Zambryski Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
description	ABSTRACT Type IV secretion systems (T4SS) transfer DNA and/or proteins into recipient cells. Here we performed immunofluorescence deconvolution microscopy to localize the assembled T4SS by detection of its native components VirB1, VirB2, VirB4, VirB5, VirB7, VirB8, VirB9, VirB10, and VirB11 in the C58 nopaline strain of Agrobacterium tumefaciens, following induction of virulence (vir) gene expression. These different proteins represent T4SS components spanning the inner membrane, periplasm, or outer membrane. Native VirB2, VirB5, VirB7, and VirB8 were also localized in the A. tumefaciens octopine strain A348. Quantitative analyses of the localization of all the above Vir proteins in nopaline and octopine strains revealed multiple foci in single optical sections in over 80% and 70% of the bacterial cells, respectively. Green fluorescent protein (GFP)-VirB8 expression following vir induction was used to monitor bacterial binding to live host plant cells; bacteria bind predominantly along their lengths, with few bacteria binding via their poles or subpoles. vir-induced attachment-defective bacteria or bacteria without the Ti plasmid do not bind to plant cells. These data support a model where multiple vir-T4SS around the perimeter of the bacterium maximize effective contact with the host to facilitate efficient transfer of DNA and protein substrates. IMPORTANCE Transfer of DNA and/or proteins to host cells through multiprotein type IV secretion system (T4SS) complexes that span the bacterial cell envelope is critical to bacterial pathogenesis. Early reports suggested that T4SS components localized at the cell poles. Now, higher-resolution deconvolution fluorescence microscopy reveals that all structural components of the Agrobacterium tumefaciens vir-T4SS, as well as its transported protein substrates, localize to multiple foci around the cell perimeter. These results lead to a new model of A. tumefaciens attachment to a plant cell, where A. tumefaciens takes advantage of the multiple vir-T4SS along its length to make intimate lateral contact with plant cells and thereby effectively transfer DNA and/or proteins through the vir-T4SS. The T4SS of A. tumefaciens is among the best-studied T4SS, and the majority of its components are highly conserved in different pathogenic bacterial species. Thus, the results presented can be applied to a broad range of pathogens that utilize T4SS.
format	article
author	Julieta Aguilar Todd A. Cameron John Zupan Patricia Zambryski
author_facet	Julieta Aguilar Todd A. Cameron John Zupan Patricia Zambryski
author_sort	Julieta Aguilar
title	Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
title_short	Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
title_full	Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
title_fullStr	Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
title_full_unstemmed	Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells
title_sort	membrane and core periplasmic <named-content content-type="genus-species">agrobacterium tumefaciens</named-content> virulence type iv secretion system components localize to multiple sites around the bacterial perimeter during lateral attachment to plant cells
publisher	American Society for Microbiology
publishDate	2011
url	https://doaj.org/article/1d25c74eda8f4a6e873bc9aa4895eb5e
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Membrane and Core Periplasmic <named-content content-type="genus-species">Agrobacterium tumefaciens</named-content> Virulence Type IV Secretion System Components Localize to Multiple Sites around the Bacterial Perimeter during Lateral Attachment to Plant Cells

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