Identification of a VapA virulence factor functional homolog in Rhodococcus equi isolates housing the pVAPB plasmid
Rhodococcus equi is a facultative intracellular bacterium of macrophages and is an important pathogen of animals and immunocompromised people wherein disease results in abcessation of the lungs and other sites. Prior work has shown that the presence of the major virulence determinant, VapA, encoded...
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| description | Rhodococcus equi is a facultative intracellular bacterium of macrophages and is an important pathogen of animals and immunocompromised people wherein disease results in abcessation of the lungs and other sites. Prior work has shown that the presence of the major virulence determinant, VapA, encoded on the pVAPA-type plasmid, disrupts normal phagosome development and is essential for bacterial replication within macrophages. pVAPA- type plasmids are typical of R. equi strains derived from foals while strains from pigs carry plasmids of the pVAPB-type, lacking vapA, and those from humans harbor various types of plasmids including pVAPA and pVAPB. Through the creation and analysis of a series of gene deletion mutants, we found that vapK1 or vapK2 is required for optimal intracellular replication of an R. equi isolate carrying a pVAPB plasmid type. Complementation analysis of a ΔvapA R. equi strain with vapK1 or vapK2 showed the VapK proteins of the pVAPB-type plasmid could restore replication capacity to the macrophage growth-attenuated ΔvapA strain. Additionally, in contrast to the intracellular growth capabilities displayed by an equine R. equi transconjugant strain carrying a pVAPB-type plasmid, a transconjugant strain carrying a pVAPB-type plasmid deleted of vapK1 and vapK2 proved incapable of replication in equine macrophages. Cumulatively, these data indicate that VapK1 and K2 are functionally equivalent to VapA. |
| doi_str_mv | 10.1371/journal.pone.0204475 |
| format | Article |
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Prior work has shown that the presence of the major virulence determinant, VapA, encoded on the pVAPA-type plasmid, disrupts normal phagosome development and is essential for bacterial replication within macrophages. pVAPA- type plasmids are typical of R. equi strains derived from foals while strains from pigs carry plasmids of the pVAPB-type, lacking vapA, and those from humans harbor various types of plasmids including pVAPA and pVAPB. Through the creation and analysis of a series of gene deletion mutants, we found that vapK1 or vapK2 is required for optimal intracellular replication of an R. equi isolate carrying a pVAPB plasmid type. Complementation analysis of a ΔvapA R. equi strain with vapK1 or vapK2 showed the VapK proteins of the pVAPB-type plasmid could restore replication capacity to the macrophage growth-attenuated ΔvapA strain. Additionally, in contrast to the intracellular growth capabilities displayed by an equine R. equi transconjugant strain carrying a pVAPB-type plasmid, a transconjugant strain carrying a pVAPB-type plasmid deleted of vapK1 and vapK2 proved incapable of replication in equine macrophages. Cumulatively, these data indicate that VapK1 and K2 are functionally equivalent to VapA.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0204475</identifier><identifier>PMID: 30286098</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Antigens ; Bacteria ; Biology and Life Sciences ; Cattle ; Clonal deletion ; Complementation ; Deletion mutant ; Evolution & development ; Gene amplification ; Gene deletion ; Genetic aspects ; Gram-positive bacteria ; Homology ; Housing ; Infections ; Infectious diseases ; Intracellular ; Juveniles ; Lungs ; Macrophages ; Medicine and Health Sciences ; Mutants ; Pathogenesis ; Physical sciences ; Physiological aspects ; Plasmids ; Proteins ; Replication ; Research and Analysis Methods ; Rhodococcus ; Rhodococcus equi ; Strains (organisms) ; Swine ; Virulence ; Virulence (Microbiology) ; Virulence factors</subject><ispartof>PloS one, 2018-10, Vol.13 (10), p.e0204475-e0204475</ispartof><rights>COPYRIGHT 2018 Public Library of Science</rights><rights>2018 Willingham-Lane et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Willingham-Lane et al 2018 Willingham-Lane et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-9e03efe3388a150f302fed10203a7800b4c9850ecc1e4bbb7874ed410e99a0693</citedby><cites>FETCH-LOGICAL-c758t-9e03efe3388a150f302fed10203a7800b4c9850ecc1e4bbb7874ed410e99a0693</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171844/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171844/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30286098$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Baruffini, Enrico</contributor><creatorcontrib>Willingham-Lane, Jennifer M</creatorcontrib><creatorcontrib>Coulson, Garry B</creatorcontrib><creatorcontrib>Hondalus, Mary K</creatorcontrib><title>Identification of a VapA virulence factor functional homolog in Rhodococcus equi isolates housing the pVAPB plasmid</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Rhodococcus equi is a facultative intracellular bacterium of macrophages and is an important pathogen of animals and immunocompromised people wherein disease results in abcessation of the lungs and other sites. Prior work has shown that the presence of the major virulence determinant, VapA, encoded on the pVAPA-type plasmid, disrupts normal phagosome development and is essential for bacterial replication within macrophages. pVAPA- type plasmids are typical of R. equi strains derived from foals while strains from pigs carry plasmids of the pVAPB-type, lacking vapA, and those from humans harbor various types of plasmids including pVAPA and pVAPB. Through the creation and analysis of a series of gene deletion mutants, we found that vapK1 or vapK2 is required for optimal intracellular replication of an R. equi isolate carrying a pVAPB plasmid type. Complementation analysis of a ΔvapA R. equi strain with vapK1 or vapK2 showed the VapK proteins of the pVAPB-type plasmid could restore replication capacity to the macrophage growth-attenuated ΔvapA strain. Additionally, in contrast to the intracellular growth capabilities displayed by an equine R. equi transconjugant strain carrying a pVAPB-type plasmid, a transconjugant strain carrying a pVAPB-type plasmid deleted of vapK1 and vapK2 proved incapable of replication in equine macrophages. Cumulatively, these data indicate that VapK1 and K2 are functionally equivalent to VapA.</description><subject>Antigens</subject><subject>Bacteria</subject><subject>Biology and Life Sciences</subject><subject>Cattle</subject><subject>Clonal deletion</subject><subject>Complementation</subject><subject>Deletion mutant</subject><subject>Evolution & development</subject><subject>Gene amplification</subject><subject>Gene deletion</subject><subject>Genetic aspects</subject><subject>Gram-positive bacteria</subject><subject>Homology</subject><subject>Housing</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>Intracellular</subject><subject>Juveniles</subject><subject>Lungs</subject><subject>Macrophages</subject><subject>Medicine and Health Sciences</subject><subject>Mutants</subject><subject>Pathogenesis</subject><subject>Physical sciences</subject><subject>Physiological 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One</addtitle><date>2018-10-04</date><risdate>2018</risdate><volume>13</volume><issue>10</issue><spage>e0204475</spage><epage>e0204475</epage><pages>e0204475-e0204475</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Rhodococcus equi is a facultative intracellular bacterium of macrophages and is an important pathogen of animals and immunocompromised people wherein disease results in abcessation of the lungs and other sites. Prior work has shown that the presence of the major virulence determinant, VapA, encoded on the pVAPA-type plasmid, disrupts normal phagosome development and is essential for bacterial replication within macrophages. pVAPA- type plasmids are typical of R. equi strains derived from foals while strains from pigs carry plasmids of the pVAPB-type, lacking vapA, and those from humans harbor various types of plasmids including pVAPA and pVAPB. Through the creation and analysis of a series of gene deletion mutants, we found that vapK1 or vapK2 is required for optimal intracellular replication of an R. equi isolate carrying a pVAPB plasmid type. Complementation analysis of a ΔvapA R. equi strain with vapK1 or vapK2 showed the VapK proteins of the pVAPB-type plasmid could restore replication capacity to the macrophage growth-attenuated ΔvapA strain. Additionally, in contrast to the intracellular growth capabilities displayed by an equine R. equi transconjugant strain carrying a pVAPB-type plasmid, a transconjugant strain carrying a pVAPB-type plasmid deleted of vapK1 and vapK2 proved incapable of replication in equine macrophages. Cumulatively, these data indicate that VapK1 and K2 are functionally equivalent to VapA.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>30286098</pmid><doi>10.1371/journal.pone.0204475</doi><tpages>e0204475</tpages><oa>free_for_read</oa></addata></record> |
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| source | DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) Journals Open Acc |
| subjects | Antigens Bacteria Biology and Life Sciences Cattle Clonal deletion Complementation Deletion mutant Evolution & development Gene amplification Gene deletion Genetic aspects Gram-positive bacteria Homology Housing Infections Infectious diseases Intracellular Juveniles Lungs Macrophages Medicine and Health Sciences Mutants Pathogenesis Physical sciences Physiological aspects Plasmids Proteins Replication Research and Analysis Methods Rhodococcus Rhodococcus equi Strains (organisms) Swine Virulence Virulence (Microbiology) Virulence factors |
| title | Identification of a VapA virulence factor functional homolog in Rhodococcus equi isolates housing the pVAPB plasmid |
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