Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells
Type IV secretion systems (T4SS) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen Agrobacterium tumefaciens uses a T4SS to deliver a VirD2‐single stranded DNA complex as well as the virulence proteins VirD5, VirE2, VirE3, and VirF into host cells so t...
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| description | Type IV secretion systems (T4SS) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen Agrobacterium tumefaciens uses a T4SS to deliver a VirD2‐single stranded DNA complex as well as the virulence proteins VirD5, VirE2, VirE3, and VirF into host cells so that these become genetically transformed. Besides plant cells, yeast and fungi can efficiently be transformed by Agrobacterium. Translocation of virulence proteins by the T4SS has so far only been shown indirectly by genetic approaches. Here we report the direct visualization of VirE2 protein translocation by using bimolecular fluorescence complementation (BiFC) and Split GFP visualization strategies. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part, either fused to VirE2 (for BiFC) or not (Split GFP). Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation. Evidence was obtained that filament formation was due to the association of VirE2 with the microtubuli.
Here we report the direct visualization of VirE2 protein translocation from Agrobacterium into host cells. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part. Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation. |
| doi_str_mv | 10.1002/mbo3.152 |
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Here we report the direct visualization of VirE2 protein translocation from Agrobacterium into host cells. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part. Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation.</description><identifier>ISSN: 2045-8827</identifier><identifier>EISSN: 2045-8827</identifier><identifier>DOI: 10.1002/mbo3.152</identifier><identifier>PMID: 24376037</identifier><language>eng</language><publisher>England: John Wiley & Sons Ltd</publisher><subject>Agrobacterium tumefaciens ; Agrobacterium tumefaciens - physiology ; Agrobacterium tumefaciens - ultrastructure ; Arabidopsis - microbiology ; Arabidopsis - ultrastructure ; Bacterial Proteins - analysis ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Bacterial Proteins - ultrastructure ; Bacterial Secretion Systems - physiology ; BiFC ; Computer Systems ; DNA-Binding Proteins - metabolism ; DNA-Binding Proteins - ultrastructure ; Flow Cytometry ; Fluorescence Resonance Energy Transfer ; Fluorescent Dyes ; Green Fluorescent Proteins - analysis ; Green Fluorescent Proteins - genetics ; Ion Channels - metabolism ; Ion Channels - ultrastructure ; Luminescent Proteins - analysis ; Luminescent Proteins - genetics ; Microscopy, Confocal ; Microtubules - microbiology ; Microtubules - physiology ; Nicotiana - microbiology ; Nicotiana - ultrastructure ; Original Research ; Peptide Fragments - analysis ; Peptide Fragments - genetics ; Protein Binding ; Protein Interaction Mapping ; protein translocation ; Protein Transport ; Protoplasts ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - ultrastructure ; split GFP ; type 4 secretion system ; VirE2</subject><ispartof>MicrobiologyOpen (Weinheim), 2014-02, Vol.3 (1), p.104-117</ispartof><rights>2013 The Authors. published by John Wiley & Sons Ltd.</rights><rights>2013 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.</rights><rights>2013 The Authors. published by John Wiley & Sons Ltd. 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937733/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937733/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,1417,11562,27924,27925,45574,45575,46052,46476,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24376037$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sakalis, Philippe A.</creatorcontrib><creatorcontrib>Heusden, G. Paul H.</creatorcontrib><creatorcontrib>Hooykaas, Paul J. J.</creatorcontrib><title>Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells</title><title>MicrobiologyOpen (Weinheim)</title><addtitle>Microbiologyopen</addtitle><description>Type IV secretion systems (T4SS) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen Agrobacterium tumefaciens uses a T4SS to deliver a VirD2‐single stranded DNA complex as well as the virulence proteins VirD5, VirE2, VirE3, and VirF into host cells so that these become genetically transformed. Besides plant cells, yeast and fungi can efficiently be transformed by Agrobacterium. Translocation of virulence proteins by the T4SS has so far only been shown indirectly by genetic approaches. Here we report the direct visualization of VirE2 protein translocation by using bimolecular fluorescence complementation (BiFC) and Split GFP visualization strategies. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part, either fused to VirE2 (for BiFC) or not (Split GFP). Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation. Evidence was obtained that filament formation was due to the association of VirE2 with the microtubuli.
Here we report the direct visualization of VirE2 protein translocation from Agrobacterium into host cells. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part. Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation.</description><subject>Agrobacterium tumefaciens</subject><subject>Agrobacterium tumefaciens - physiology</subject><subject>Agrobacterium tumefaciens - ultrastructure</subject><subject>Arabidopsis - microbiology</subject><subject>Arabidopsis - ultrastructure</subject><subject>Bacterial Proteins - analysis</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacterial Proteins - ultrastructure</subject><subject>Bacterial Secretion Systems - physiology</subject><subject>BiFC</subject><subject>Computer Systems</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>DNA-Binding Proteins - ultrastructure</subject><subject>Flow Cytometry</subject><subject>Fluorescence Resonance Energy Transfer</subject><subject>Fluorescent Dyes</subject><subject>Green Fluorescent Proteins - analysis</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Ion Channels - metabolism</subject><subject>Ion Channels - ultrastructure</subject><subject>Luminescent Proteins - analysis</subject><subject>Luminescent Proteins - genetics</subject><subject>Microscopy, Confocal</subject><subject>Microtubules - microbiology</subject><subject>Microtubules - physiology</subject><subject>Nicotiana - microbiology</subject><subject>Nicotiana - ultrastructure</subject><subject>Original Research</subject><subject>Peptide Fragments - analysis</subject><subject>Peptide Fragments - genetics</subject><subject>Protein Binding</subject><subject>Protein Interaction Mapping</subject><subject>protein translocation</subject><subject>Protein Transport</subject><subject>Protoplasts</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - ultrastructure</subject><subject>split GFP</subject><subject>type 4 secretion system</subject><subject>VirE2</subject><issn>2045-8827</issn><issn>2045-8827</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>EIF</sourceid><recordid>eNqFkU9r3DAQxUVpaEKy0E9QdMzFqf7Zki-BbUjTQMpekr0KWR5nFWzLkeQW99PH290um1PnMgPvx2MeD6HPlFxRQtjXrvL8iubsAzpjROSZUkx-PLpP0SLGFzKPJKwQ9BM6ZYLLgnB5hl7WLo6mdX9Mcr7HvsFrF24ZHoJP4Hqcgulj6-1OriacNoCXz8FXxiYIbuxwmgbA92scwQb4i8UpJuiw65PHGx8TttC28QKdNKaNsNjvc_T0_fbx5kf2sLq7v1k-ZAOXlGWsLmtrG8KUUiUnYHIJoiJgaa0syRsCczJRKcaqAoggjVB5XZu8KRTQQjJ-jq53vsNYdVBb6OcQrR6C60yYtDdOv1d6t9HP_pfmJZeS89ngcm8Q_OsIMenOxW0E04Mfo6aSSlEypsj_UVGWczVMbtEvx28d_vnXxQxkO-C3a2E66JTobc16W7OevfTPbys-b_4GAZ2bPg</recordid><startdate>201402</startdate><enddate>201402</enddate><creator>Sakalis, Philippe A.</creator><creator>Heusden, G. Paul H.</creator><creator>Hooykaas, Paul J. J.</creator><general>John Wiley & Sons Ltd</general><scope>24P</scope><scope>WIN</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>5PM</scope></search><sort><creationdate>201402</creationdate><title>Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells</title><author>Sakalis, Philippe A. ; Heusden, G. Paul H. ; Hooykaas, Paul J. J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p3712-2d9dccf02888930ea57e4b0ec1d8c05f0e2044b822b6e040f485dda5f68e16723</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Agrobacterium tumefaciens</topic><topic>Agrobacterium tumefaciens - physiology</topic><topic>Agrobacterium tumefaciens - ultrastructure</topic><topic>Arabidopsis - microbiology</topic><topic>Arabidopsis - ultrastructure</topic><topic>Bacterial Proteins - analysis</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Bacterial Proteins - ultrastructure</topic><topic>Bacterial Secretion Systems - physiology</topic><topic>BiFC</topic><topic>Computer Systems</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>DNA-Binding Proteins - ultrastructure</topic><topic>Flow Cytometry</topic><topic>Fluorescence Resonance Energy Transfer</topic><topic>Fluorescent Dyes</topic><topic>Green Fluorescent Proteins - analysis</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Ion Channels - metabolism</topic><topic>Ion Channels - ultrastructure</topic><topic>Luminescent Proteins - analysis</topic><topic>Luminescent Proteins - genetics</topic><topic>Microscopy, Confocal</topic><topic>Microtubules - microbiology</topic><topic>Microtubules - physiology</topic><topic>Nicotiana - microbiology</topic><topic>Nicotiana - ultrastructure</topic><topic>Original Research</topic><topic>Peptide Fragments - analysis</topic><topic>Peptide Fragments - genetics</topic><topic>Protein Binding</topic><topic>Protein Interaction Mapping</topic><topic>protein translocation</topic><topic>Protein Transport</topic><topic>Protoplasts</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - ultrastructure</topic><topic>split GFP</topic><topic>type 4 secretion system</topic><topic>VirE2</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sakalis, Philippe A.</creatorcontrib><creatorcontrib>Heusden, G. Paul H.</creatorcontrib><creatorcontrib>Hooykaas, Paul J. J.</creatorcontrib><collection>Wiley-Blackwell Open Access Titles</collection><collection>Wiley Free Content</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>MicrobiologyOpen (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sakalis, Philippe A.</au><au>Heusden, G. Paul H.</au><au>Hooykaas, Paul J. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells</atitle><jtitle>MicrobiologyOpen (Weinheim)</jtitle><addtitle>Microbiologyopen</addtitle><date>2014-02</date><risdate>2014</risdate><volume>3</volume><issue>1</issue><spage>104</spage><epage>117</epage><pages>104-117</pages><issn>2045-8827</issn><eissn>2045-8827</eissn><abstract>Type IV secretion systems (T4SS) can mediate the translocation of bacterial virulence proteins into host cells. The plant pathogen Agrobacterium tumefaciens uses a T4SS to deliver a VirD2‐single stranded DNA complex as well as the virulence proteins VirD5, VirE2, VirE3, and VirF into host cells so that these become genetically transformed. Besides plant cells, yeast and fungi can efficiently be transformed by Agrobacterium. Translocation of virulence proteins by the T4SS has so far only been shown indirectly by genetic approaches. Here we report the direct visualization of VirE2 protein translocation by using bimolecular fluorescence complementation (BiFC) and Split GFP visualization strategies. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part, either fused to VirE2 (for BiFC) or not (Split GFP). Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation. Evidence was obtained that filament formation was due to the association of VirE2 with the microtubuli.
Here we report the direct visualization of VirE2 protein translocation from Agrobacterium into host cells. To this end, we cocultivated Agrobacterium strains expressing VirE2 tagged with one part of a fluorescent protein with host cells expressing the complementary part. Fluorescent filaments became visible in recipient cells 20–25 h after the start of the cocultivation indicative of VirE2 protein translocation.</abstract><cop>England</cop><pub>John Wiley & Sons Ltd</pub><pmid>24376037</pmid><doi>10.1002/mbo3.152</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Agrobacterium tumefaciens Agrobacterium tumefaciens - physiology Agrobacterium tumefaciens - ultrastructure Arabidopsis - microbiology Arabidopsis - ultrastructure Bacterial Proteins - analysis Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacterial Proteins - ultrastructure Bacterial Secretion Systems - physiology BiFC Computer Systems DNA-Binding Proteins - metabolism DNA-Binding Proteins - ultrastructure Flow Cytometry Fluorescence Resonance Energy Transfer Fluorescent Dyes Green Fluorescent Proteins - analysis Green Fluorescent Proteins - genetics Ion Channels - metabolism Ion Channels - ultrastructure Luminescent Proteins - analysis Luminescent Proteins - genetics Microscopy, Confocal Microtubules - microbiology Microtubules - physiology Nicotiana - microbiology Nicotiana - ultrastructure Original Research Peptide Fragments - analysis Peptide Fragments - genetics Protein Binding Protein Interaction Mapping protein translocation Protein Transport Protoplasts Saccharomyces cerevisiae Saccharomyces cerevisiae - ultrastructure split GFP type 4 secretion system VirE2 |
| title | Visualization of VirE2 protein translocation by the Agrobacterium type IV secretion system into host cells |
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