The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis
All dsDNA phages encode two proteins involved in host lysis, an endolysin and a holin that target the peptidoglycan and cytoplasmic membrane, respectively. Bacteriophages that infect Gram-negative bacteria encode additional proteins, the spanins, involved in disruption of the outer membrane. Recentl...
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MDPI AG
2017-11-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/9/11/343 |
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| author | Adriano M. Gigante Cheri M. Hampton Rebecca S. Dillard Filipa Gil Maria João Catalão José Moniz-Pereira Elizabeth R. Wright Madalena Pimentel |
| author_facet | Adriano M. Gigante Cheri M. Hampton Rebecca S. Dillard Filipa Gil Maria João Catalão José Moniz-Pereira Elizabeth R. Wright Madalena Pimentel |
| author_sort | Adriano M. Gigante |
| collection | DOAJ |
| description | All dsDNA phages encode two proteins involved in host lysis, an endolysin and a holin that target the peptidoglycan and cytoplasmic membrane, respectively. Bacteriophages that infect Gram-negative bacteria encode additional proteins, the spanins, involved in disruption of the outer membrane. Recently, a gene located in the lytic cassette was identified in the genomes of mycobacteriophages, which encodes a protein (LysB) with mycolyl-arabinogalactan esterase activity. Taking in consideration the complex mycobacterial cell envelope that mycobacteriophages encounter during their life cycle, it is valuable to evaluate the role of these proteins in lysis. In the present work, we constructed an Ms6 mutant defective on lysB and showed that Ms6 LysB has an important role in lysis. In the absence of LysB, lysis still occurs but the newly synthesized phage particles are deficiently released to the environment. Using cryo-electron microscopy and tomography to register the changes in the lysis phenotype, we show that at 150 min post-adsorption, mycobacteria cells are incompletely lysed and phage particles are retained inside the cell, while cells infected with Ms6wt are completely lysed. Our results confirm that Ms6 LysB is necessary for an efficient lysis of Mycobacterium smegmatis, acting, similarly to spanins, in the third step of the lysis process. |
| first_indexed | 2024-12-23T05:03:20Z |
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| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-23T05:03:20Z |
| publishDate | 2017-11-01 |
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| series | Viruses |
| spelling | doaj.art-4e939f1279f544e6821c17c7a92471c42022-12-21T17:59:10ZengMDPI AGViruses1999-49152017-11-0191134310.3390/v9110343v9110343The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced LysisAdriano M. Gigante0Cheri M. Hampton1Rebecca S. Dillard2Filipa Gil3Maria João Catalão4José Moniz-Pereira5Elizabeth R. Wright6Madalena Pimentel7Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, 1649-003, PortugalDivision of Pediatric Infectious Diseases, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30345, USADivision of Pediatric Infectious Diseases, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30345, USAResearch Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, 1649-003, PortugalResearch Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, 1649-003, PortugalResearch Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, 1649-003, PortugalDivision of Pediatric Infectious Diseases, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30345, USAResearch Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Lisbon, 1649-003, PortugalAll dsDNA phages encode two proteins involved in host lysis, an endolysin and a holin that target the peptidoglycan and cytoplasmic membrane, respectively. Bacteriophages that infect Gram-negative bacteria encode additional proteins, the spanins, involved in disruption of the outer membrane. Recently, a gene located in the lytic cassette was identified in the genomes of mycobacteriophages, which encodes a protein (LysB) with mycolyl-arabinogalactan esterase activity. Taking in consideration the complex mycobacterial cell envelope that mycobacteriophages encounter during their life cycle, it is valuable to evaluate the role of these proteins in lysis. In the present work, we constructed an Ms6 mutant defective on lysB and showed that Ms6 LysB has an important role in lysis. In the absence of LysB, lysis still occurs but the newly synthesized phage particles are deficiently released to the environment. Using cryo-electron microscopy and tomography to register the changes in the lysis phenotype, we show that at 150 min post-adsorption, mycobacteria cells are incompletely lysed and phage particles are retained inside the cell, while cells infected with Ms6wt are completely lysed. Our results confirm that Ms6 LysB is necessary for an efficient lysis of Mycobacterium smegmatis, acting, similarly to spanins, in the third step of the lysis process.https://www.mdpi.com/1999-4915/9/11/343bacteriophage lysismycobacteriophageMs6LysBmycobacteriaspaninscryo-electron microscopy |
| spellingShingle | Adriano M. Gigante Cheri M. Hampton Rebecca S. Dillard Filipa Gil Maria João Catalão José Moniz-Pereira Elizabeth R. Wright Madalena Pimentel The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis Viruses bacteriophage lysis mycobacteriophage Ms6 LysB mycobacteria spanins cryo-electron microscopy |
| title | The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis |
| title_full | The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis |
| title_fullStr | The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis |
| title_full_unstemmed | The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis |
| title_short | The Ms6 Mycolyl-Arabinogalactan Esterase LysB is Essential for an Efficient Mycobacteriophage-Induced Lysis |
| title_sort | ms6 mycolyl arabinogalactan esterase lysb is essential for an efficient mycobacteriophage induced lysis |
| topic | bacteriophage lysis mycobacteriophage Ms6 LysB mycobacteria spanins cryo-electron microscopy |
| url | https://www.mdpi.com/1999-4915/9/11/343 |
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