Analysis of SecA2‐dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector
Summary The pathogenicity of mycobacteria is closely associated with their ability to export virulence factors. For this purpose, mycobacteria possess different protein secretion systems, including the accessory Sec translocation pathway, SecA2. Although this pathway is associated with intracellular...
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| Veröffentlicht in: | Cellular microbiology 2014-02, Vol.16 (2), p.280-295 |
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| creator | Woude, Aniek D. Stoop, Esther J. M. Stiess, Michael Wang, Sen Ummels, Roy Stempvoort, Gunny Piersma, Sander R. Cascioferro, Alessandro Jiménez, Connie R. Houben, Edith N. G. Luirink, Joen Pieters, Jean Sar, Astrid M. Bitter, Wilbert |
| description | Summary
The pathogenicity of mycobacteria is closely associated with their ability to export virulence factors. For this purpose, mycobacteria possess different protein secretion systems, including the accessory Sec translocation pathway, SecA2. Although this pathway is associated with intracellular survival and virulence, the SecA2‐dependent effector proteins remain largely undefined. In this work, we studied a Mycobacterium marinum secA2 mutant with an impaired capacity to initiate granuloma formation in zebrafish embryos. By comparing the proteomic profile of cell envelope fractions from the secA2 mutant with wild type M. marinum, we identified putative SecA2‐dependent substrates. Immunoblotting procedures confirmed SecA2‐dependent membrane localization for several of these proteins, including the virulence factor protein kinase G (PknG). Interestingly, phenotypical defects of the secA2 mutant are similar to those described for ΔpknG, including phagosomal maturation. Overexpression of PknG in the secA2 mutant restored its localization to the cell envelope. Importantly, PknG‐overexpression also partially restored the virulence of the secA2 mutant, as indicated by enhanced infectivity in zebrafish embryos and restored inhibition of phagosomal maturation. These results suggest that SecA2‐dependent membrane localization of PknG is an important determinant for M. marinum virulence. |
| doi_str_mv | 10.1111/cmi.12221 |
| format | Article |
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The pathogenicity of mycobacteria is closely associated with their ability to export virulence factors. For this purpose, mycobacteria possess different protein secretion systems, including the accessory Sec translocation pathway, SecA2. Although this pathway is associated with intracellular survival and virulence, the SecA2‐dependent effector proteins remain largely undefined. In this work, we studied a Mycobacterium marinum secA2 mutant with an impaired capacity to initiate granuloma formation in zebrafish embryos. By comparing the proteomic profile of cell envelope fractions from the secA2 mutant with wild type M. marinum, we identified putative SecA2‐dependent substrates. Immunoblotting procedures confirmed SecA2‐dependent membrane localization for several of these proteins, including the virulence factor protein kinase G (PknG). Interestingly, phenotypical defects of the secA2 mutant are similar to those described for ΔpknG, including phagosomal maturation. Overexpression of PknG in the secA2 mutant restored its localization to the cell envelope. Importantly, PknG‐overexpression also partially restored the virulence of the secA2 mutant, as indicated by enhanced infectivity in zebrafish embryos and restored inhibition of phagosomal maturation. These results suggest that SecA2‐dependent membrane localization of PknG is an important determinant for M. marinum virulence.</description><identifier>ISSN: 1462-5814</identifier><identifier>EISSN: 1462-5822</identifier><identifier>DOI: 10.1111/cmi.12221</identifier><identifier>PMID: 24119166</identifier><language>eng</language><publisher>England: Hindawi Limited</publisher><subject>Adenosine Triphosphatases - metabolism ; Animals ; Bacterial Proteins - metabolism ; Bacteriology ; Cyclic GMP-Dependent Protein Kinases - metabolism ; Danio rerio ; Disease Models, Animal ; DNA Transposable Elements ; Gene Knockout Techniques ; Immunoblotting ; Kinases ; Membrane Transport Proteins - metabolism ; Mutagenesis, Insertional ; Mycobacterium ; Mycobacterium Infections - microbiology ; Mycobacterium marinum ; Mycobacterium marinum - metabolism ; Mycobacterium marinum - pathogenicity ; Proteins ; Substrate Specificity ; Virulence Factors - metabolism ; Zebrafish</subject><ispartof>Cellular microbiology, 2014-02, Vol.16 (2), p.280-295</ispartof><rights>2013 John Wiley & Sons Ltd</rights><rights>2013 John Wiley & Sons Ltd.</rights><rights>Copyright © 2014 John Wiley & Sons Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fcmi.12221$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fcmi.12221$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24119166$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Woude, Aniek D.</creatorcontrib><creatorcontrib>Stoop, Esther J. M.</creatorcontrib><creatorcontrib>Stiess, Michael</creatorcontrib><creatorcontrib>Wang, Sen</creatorcontrib><creatorcontrib>Ummels, Roy</creatorcontrib><creatorcontrib>Stempvoort, Gunny</creatorcontrib><creatorcontrib>Piersma, Sander R.</creatorcontrib><creatorcontrib>Cascioferro, Alessandro</creatorcontrib><creatorcontrib>Jiménez, Connie R.</creatorcontrib><creatorcontrib>Houben, Edith N. G.</creatorcontrib><creatorcontrib>Luirink, Joen</creatorcontrib><creatorcontrib>Pieters, Jean</creatorcontrib><creatorcontrib>Sar, Astrid M.</creatorcontrib><creatorcontrib>Bitter, Wilbert</creatorcontrib><title>Analysis of SecA2‐dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector</title><title>Cellular microbiology</title><addtitle>Cell Microbiol</addtitle><description>Summary
The pathogenicity of mycobacteria is closely associated with their ability to export virulence factors. For this purpose, mycobacteria possess different protein secretion systems, including the accessory Sec translocation pathway, SecA2. Although this pathway is associated with intracellular survival and virulence, the SecA2‐dependent effector proteins remain largely undefined. In this work, we studied a Mycobacterium marinum secA2 mutant with an impaired capacity to initiate granuloma formation in zebrafish embryos. By comparing the proteomic profile of cell envelope fractions from the secA2 mutant with wild type M. marinum, we identified putative SecA2‐dependent substrates. Immunoblotting procedures confirmed SecA2‐dependent membrane localization for several of these proteins, including the virulence factor protein kinase G (PknG). Interestingly, phenotypical defects of the secA2 mutant are similar to those described for ΔpknG, including phagosomal maturation. Overexpression of PknG in the secA2 mutant restored its localization to the cell envelope. Importantly, PknG‐overexpression also partially restored the virulence of the secA2 mutant, as indicated by enhanced infectivity in zebrafish embryos and restored inhibition of phagosomal maturation. These results suggest that SecA2‐dependent membrane localization of PknG is an important determinant for M. marinum virulence.</description><subject>Adenosine Triphosphatases - metabolism</subject><subject>Animals</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacteriology</subject><subject>Cyclic GMP-Dependent Protein Kinases - metabolism</subject><subject>Danio rerio</subject><subject>Disease Models, Animal</subject><subject>DNA Transposable Elements</subject><subject>Gene Knockout Techniques</subject><subject>Immunoblotting</subject><subject>Kinases</subject><subject>Membrane Transport Proteins - metabolism</subject><subject>Mutagenesis, Insertional</subject><subject>Mycobacterium</subject><subject>Mycobacterium Infections - microbiology</subject><subject>Mycobacterium marinum</subject><subject>Mycobacterium marinum - metabolism</subject><subject>Mycobacterium marinum - pathogenicity</subject><subject>Proteins</subject><subject>Substrate Specificity</subject><subject>Virulence Factors - metabolism</subject><subject>Zebrafish</subject><issn>1462-5814</issn><issn>1462-5822</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0U9LwzAYBvAgipt_Dn4BCXjRw1zfpEnb4xg6BYeCei5p-haytelsWmXgwY_gZ_STGLe5gydzeQP58RKeh5ATCC7Bn6GuzCUwxmCH9CGUbCBixna3dwh75MC5WRCAjAD2SY-FAAlI2SfvI6vKpTOO1gV9RD1iXx-fOS7Q5mhb6rrMtY1q0VFj6XSp60zpFhvTVbRSjbF-mh9pCuPNoqlb9HBurHJIJ_T8YW4nF1Q5quiraboSrUaKRYG6rZsjsleo0uHxZh6S5-urp_HN4O5-cjse3Q0WHBgMIi1DkLnkBculDBMUhcxklOQ841LEMQSBTlSQIItFnKHQTCjFlBAqDGO_gB-S8_Ve_7-XDl2bVsZpLEtlse5cCoIx6WPi_6GB4CzyoXp69ofO6q7xcXoVRjIRIk64V6cb1WUV5umiMT64ZfpbgQfDNXgzJS637xCkP92mvtt01W06nt6uLvwbteGVTQ</recordid><startdate>201402</startdate><enddate>201402</enddate><creator>Woude, Aniek D.</creator><creator>Stoop, Esther J. M.</creator><creator>Stiess, Michael</creator><creator>Wang, Sen</creator><creator>Ummels, Roy</creator><creator>Stempvoort, Gunny</creator><creator>Piersma, Sander R.</creator><creator>Cascioferro, Alessandro</creator><creator>Jiménez, Connie R.</creator><creator>Houben, Edith N. G.</creator><creator>Luirink, Joen</creator><creator>Pieters, Jean</creator><creator>Sar, Astrid M.</creator><creator>Bitter, Wilbert</creator><general>Hindawi Limited</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QL</scope><scope>7T7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201402</creationdate><title>Analysis of SecA2‐dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector</title><author>Woude, Aniek D. ; Stoop, Esther J. M. ; Stiess, Michael ; Wang, Sen ; Ummels, Roy ; Stempvoort, Gunny ; Piersma, Sander R. ; Cascioferro, Alessandro ; Jiménez, Connie R. ; Houben, Edith N. 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M.</creatorcontrib><creatorcontrib>Stiess, Michael</creatorcontrib><creatorcontrib>Wang, Sen</creatorcontrib><creatorcontrib>Ummels, Roy</creatorcontrib><creatorcontrib>Stempvoort, Gunny</creatorcontrib><creatorcontrib>Piersma, Sander R.</creatorcontrib><creatorcontrib>Cascioferro, Alessandro</creatorcontrib><creatorcontrib>Jiménez, Connie R.</creatorcontrib><creatorcontrib>Houben, Edith N. G.</creatorcontrib><creatorcontrib>Luirink, Joen</creatorcontrib><creatorcontrib>Pieters, Jean</creatorcontrib><creatorcontrib>Sar, Astrid M.</creatorcontrib><creatorcontrib>Bitter, Wilbert</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cellular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Woude, Aniek D.</au><au>Stoop, Esther J. M.</au><au>Stiess, Michael</au><au>Wang, Sen</au><au>Ummels, Roy</au><au>Stempvoort, Gunny</au><au>Piersma, Sander R.</au><au>Cascioferro, Alessandro</au><au>Jiménez, Connie R.</au><au>Houben, Edith N. G.</au><au>Luirink, Joen</au><au>Pieters, Jean</au><au>Sar, Astrid M.</au><au>Bitter, Wilbert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analysis of SecA2‐dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector</atitle><jtitle>Cellular microbiology</jtitle><addtitle>Cell Microbiol</addtitle><date>2014-02</date><risdate>2014</risdate><volume>16</volume><issue>2</issue><spage>280</spage><epage>295</epage><pages>280-295</pages><issn>1462-5814</issn><eissn>1462-5822</eissn><abstract>Summary
The pathogenicity of mycobacteria is closely associated with their ability to export virulence factors. For this purpose, mycobacteria possess different protein secretion systems, including the accessory Sec translocation pathway, SecA2. Although this pathway is associated with intracellular survival and virulence, the SecA2‐dependent effector proteins remain largely undefined. In this work, we studied a Mycobacterium marinum secA2 mutant with an impaired capacity to initiate granuloma formation in zebrafish embryos. By comparing the proteomic profile of cell envelope fractions from the secA2 mutant with wild type M. marinum, we identified putative SecA2‐dependent substrates. Immunoblotting procedures confirmed SecA2‐dependent membrane localization for several of these proteins, including the virulence factor protein kinase G (PknG). Interestingly, phenotypical defects of the secA2 mutant are similar to those described for ΔpknG, including phagosomal maturation. Overexpression of PknG in the secA2 mutant restored its localization to the cell envelope. Importantly, PknG‐overexpression also partially restored the virulence of the secA2 mutant, as indicated by enhanced infectivity in zebrafish embryos and restored inhibition of phagosomal maturation. These results suggest that SecA2‐dependent membrane localization of PknG is an important determinant for M. marinum virulence.</abstract><cop>England</cop><pub>Hindawi Limited</pub><pmid>24119166</pmid><doi>10.1111/cmi.12221</doi><tpages>16</tpages></addata></record> |
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| subjects | Adenosine Triphosphatases - metabolism Animals Bacterial Proteins - metabolism Bacteriology Cyclic GMP-Dependent Protein Kinases - metabolism Danio rerio Disease Models, Animal DNA Transposable Elements Gene Knockout Techniques Immunoblotting Kinases Membrane Transport Proteins - metabolism Mutagenesis, Insertional Mycobacterium Mycobacterium Infections - microbiology Mycobacterium marinum Mycobacterium marinum - metabolism Mycobacterium marinum - pathogenicity Proteins Substrate Specificity Virulence Factors - metabolism Zebrafish |
| title | Analysis of SecA2‐dependent substrates in Mycobacterium marinum identifies protein kinase G (PknG) as a virulence effector |
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