The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i>
<i>Klebsiella pneumoniae</i> is considered one of the most critical multidrug-resistant pathogens and urgently requires new therapeutic strategies. Capsular polysaccharides (CPS), lipopolysaccharides (LPS), and exopolysaccharides (EPS) are the major virulence factors protecting <i>...
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MDPI AG
2021-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/21/11562 |
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| author | Marta Kaszowska Grazyna Majkowska-Skrobek Pawel Markwitz Cédric Lood Wojciech Jachymek Anna Maciejewska Jolanta Lukasiewicz Zuzanna Drulis-Kawa |
| author_facet | Marta Kaszowska Grazyna Majkowska-Skrobek Pawel Markwitz Cédric Lood Wojciech Jachymek Anna Maciejewska Jolanta Lukasiewicz Zuzanna Drulis-Kawa |
| author_sort | Marta Kaszowska |
| collection | DOAJ |
| description | <i>Klebsiella pneumoniae</i> is considered one of the most critical multidrug-resistant pathogens and urgently requires new therapeutic strategies. Capsular polysaccharides (CPS), lipopolysaccharides (LPS), and exopolysaccharides (EPS) are the major virulence factors protecting <i>K. pneumoniae</i> against the immune response and thus may be targeted by phage-based therapeutics such as polysaccharides-degrading enzymes. Since the emergence of resistance to antibacterials is generally considered undesirable, in this study, the genetic and phenotypic characteristics of resistance to the phage-borne CPS-degrading depolymerase and its effect on <i>K. pneumoniae</i> virulence were investigated. The K63 serotype targeting depolymerase (KP36gp50) derived from <i>Klebsiella</i> siphovirus KP36 was used as the selective agent during the treatment of <i>K. pneumoniae</i> 486 biofilm. Genome-driven examination combined with the surface polysaccharide structural analysis of resistant mutant showed the point mutation and frameshift in the <i>wbaP</i> gene located within the <i>cps</i> gene cluster, resulting in the loss of the capsule. The sharp decline in the yield of CPS was accompanied by the production of a larger amount of smooth LPS. The modification of the surface polysaccharide layers did not affect bacterial fitness nor the insensitivity to serum complement; however, it made bacteria more prone to phagocytosis combined with the higher adherence and internalization to human lung epithelial cells. In that context, it was showed that the emerging resistance to the antivirulence agent (phage-borne capsule depolymerase) results in beneficial consequences, i.e., the sensitization to the innate immune response. |
| first_indexed | 2024-03-10T06:02:08Z |
| format | Article |
| id | doaj.art-e4b8bc86bdb742698ded9020d437732a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T06:02:08Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e4b8bc86bdb742698ded9020d437732a2023-11-22T20:54:13ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-10-0122211156210.3390/ijms222111562The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i>Marta Kaszowska0Grazyna Majkowska-Skrobek1Pawel Markwitz2Cédric Lood3Wojciech Jachymek4Anna Maciejewska5Jolanta Lukasiewicz6Zuzanna Drulis-Kawa7Laboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53-114 Wroclaw, PolandDepartment of Pathogen Biology and Immunology, University of Wroclaw, 51-148 Wroclaw, PolandDepartment of Pathogen Biology and Immunology, University of Wroclaw, 51-148 Wroclaw, PolandDepartment of Microbial and Molecular Systems, KU Leuven, 3001 Leuven, BelgiumLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53-114 Wroclaw, PolandLaboratory of Microbial Immunochemistry and Vaccines, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53-114 Wroclaw, PolandDepartment of Pathogen Biology and Immunology, University of Wroclaw, 51-148 Wroclaw, Poland<i>Klebsiella pneumoniae</i> is considered one of the most critical multidrug-resistant pathogens and urgently requires new therapeutic strategies. Capsular polysaccharides (CPS), lipopolysaccharides (LPS), and exopolysaccharides (EPS) are the major virulence factors protecting <i>K. pneumoniae</i> against the immune response and thus may be targeted by phage-based therapeutics such as polysaccharides-degrading enzymes. Since the emergence of resistance to antibacterials is generally considered undesirable, in this study, the genetic and phenotypic characteristics of resistance to the phage-borne CPS-degrading depolymerase and its effect on <i>K. pneumoniae</i> virulence were investigated. The K63 serotype targeting depolymerase (KP36gp50) derived from <i>Klebsiella</i> siphovirus KP36 was used as the selective agent during the treatment of <i>K. pneumoniae</i> 486 biofilm. Genome-driven examination combined with the surface polysaccharide structural analysis of resistant mutant showed the point mutation and frameshift in the <i>wbaP</i> gene located within the <i>cps</i> gene cluster, resulting in the loss of the capsule. The sharp decline in the yield of CPS was accompanied by the production of a larger amount of smooth LPS. The modification of the surface polysaccharide layers did not affect bacterial fitness nor the insensitivity to serum complement; however, it made bacteria more prone to phagocytosis combined with the higher adherence and internalization to human lung epithelial cells. In that context, it was showed that the emerging resistance to the antivirulence agent (phage-borne capsule depolymerase) results in beneficial consequences, i.e., the sensitization to the innate immune response.https://www.mdpi.com/1422-0067/22/21/11562<i>Klebsiella</i> phagecapsule degrading depolymerasephage resistant mutantcapsular polysaccharide |
| spellingShingle | Marta Kaszowska Grazyna Majkowska-Skrobek Pawel Markwitz Cédric Lood Wojciech Jachymek Anna Maciejewska Jolanta Lukasiewicz Zuzanna Drulis-Kawa The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> International Journal of Molecular Sciences <i>Klebsiella</i> phage capsule degrading depolymerase phage resistant mutant capsular polysaccharide |
| title | The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> |
| title_full | The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> |
| title_fullStr | The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> |
| title_full_unstemmed | The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> |
| title_short | The Mutation in <i>wbaP cps</i> Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of <i>Klebsiella pneumoniae</i> |
| title_sort | mutation in i wbap cps i gene cluster selected by phage borne depolymerase abolishes capsule production and diminishes the virulence of i klebsiella pneumoniae i |
| topic | <i>Klebsiella</i> phage capsule degrading depolymerase phage resistant mutant capsular polysaccharide |
| url | https://www.mdpi.com/1422-0067/22/21/11562 |
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