RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6
Bacteriophages (phages) are predicted to be the most ubiquitous biological entity on earth, and yet, there are still vast knowledge gaps in our understanding of phage diversity and phage–host interactions. Approximately one hundred <i>Acinetobacter</i>-infecting DNA viruses have been ide...
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MDPI AG
2021-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/8/1652 |
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| author | Clay S. Crippen Bibi Zhou Silke Andresen Robert T. Patry Artur Muszyński Craig T. Parker Kerry K. Cooper Christine M. Szymanski |
| author_facet | Clay S. Crippen Bibi Zhou Silke Andresen Robert T. Patry Artur Muszyński Craig T. Parker Kerry K. Cooper Christine M. Szymanski |
| author_sort | Clay S. Crippen |
| collection | DOAJ |
| description | Bacteriophages (phages) are predicted to be the most ubiquitous biological entity on earth, and yet, there are still vast knowledge gaps in our understanding of phage diversity and phage–host interactions. Approximately one hundred <i>Acinetobacter</i>-infecting DNA viruses have been identified, and in this report, we describe eight more. We isolated two typical dsDNA lytic podoviruses (CAP1–2), five unique dsRNA lytic cystoviruses (CAP3–7), and one dsDNA lysogenic siphovirus (SLAP1), all capable of infecting the multidrug resistant isolate <i>Acinetobacter radioresistens</i> LH6. Using transmission electron microscopy, bacterial mutagenesis, phage infectivity assays, carbohydrate staining, mass-spectrometry, genomic sequencing, and comparative studies, we further characterized these phages. Mutation of the LH6 initiating glycosyltransferase homolog, PglC, necessary for both O-linked glycoprotein and capsular polysaccharide (CPS) biosynthesis, prevented infection by the lytic podovirus CAP1, while mutation of the pilin protein, PilA, prevented infection by CAP3, representing the lytic cystoviruses. Genome sequencing of the three dsRNA segments of the isolated cystoviruses revealed low levels of homology, but conserved synteny with the only other reported cystoviruses that infect <i>Pseudomonas</i> species. In <i>Pseudomonas</i>, the cystoviruses are known to be enveloped phages surrounding their capsids with the inner membrane from the infected host. To characterize any membrane-associated glycoconjugates in the CAP3 cystovirus, carbohydrate staining was used to identify a low molecular weight lipid-linked glycoconjugate subsequently identified by mutagenesis and mass-spectrometry as bacterial lipooligosaccharide. Together, this study demonstrates the isolation of new <i>Acinetobacter</i>-infecting phages and the determination of their cell receptors. Further, we describe the genomes of a new genus of <i>Cystoviruses</i> and perform an initial characterization of membrane-associated glycoconjugates. |
| first_indexed | 2024-03-10T08:17:59Z |
| format | Article |
| id | doaj.art-d148c9cb9d9e49a0b3a235fe2f8972f9 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T08:17:59Z |
| publishDate | 2021-08-01 |
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| spelling | doaj.art-d148c9cb9d9e49a0b3a235fe2f8972f92023-11-22T10:12:47ZengMDPI AGViruses1999-49152021-08-01138165210.3390/v13081652RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6Clay S. Crippen0Bibi Zhou1Silke Andresen2Robert T. Patry3Artur Muszyński4Craig T. Parker5Kerry K. Cooper6Christine M. Szymanski7Department of Microbiology, University of Georgia, Athens, GA 30602, USADepartment of Microbiology, University of Georgia, Athens, GA 30602, USADepartment of Microbiology, University of Georgia, Athens, GA 30602, USADepartment of Microbiology, University of Georgia, Athens, GA 30602, USAComplex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USAProduce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, CA 94710, USASchool of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, AZ 85721, USADepartment of Microbiology, University of Georgia, Athens, GA 30602, USABacteriophages (phages) are predicted to be the most ubiquitous biological entity on earth, and yet, there are still vast knowledge gaps in our understanding of phage diversity and phage–host interactions. Approximately one hundred <i>Acinetobacter</i>-infecting DNA viruses have been identified, and in this report, we describe eight more. We isolated two typical dsDNA lytic podoviruses (CAP1–2), five unique dsRNA lytic cystoviruses (CAP3–7), and one dsDNA lysogenic siphovirus (SLAP1), all capable of infecting the multidrug resistant isolate <i>Acinetobacter radioresistens</i> LH6. Using transmission electron microscopy, bacterial mutagenesis, phage infectivity assays, carbohydrate staining, mass-spectrometry, genomic sequencing, and comparative studies, we further characterized these phages. Mutation of the LH6 initiating glycosyltransferase homolog, PglC, necessary for both O-linked glycoprotein and capsular polysaccharide (CPS) biosynthesis, prevented infection by the lytic podovirus CAP1, while mutation of the pilin protein, PilA, prevented infection by CAP3, representing the lytic cystoviruses. Genome sequencing of the three dsRNA segments of the isolated cystoviruses revealed low levels of homology, but conserved synteny with the only other reported cystoviruses that infect <i>Pseudomonas</i> species. In <i>Pseudomonas</i>, the cystoviruses are known to be enveloped phages surrounding their capsids with the inner membrane from the infected host. To characterize any membrane-associated glycoconjugates in the CAP3 cystovirus, carbohydrate staining was used to identify a low molecular weight lipid-linked glycoconjugate subsequently identified by mutagenesis and mass-spectrometry as bacterial lipooligosaccharide. Together, this study demonstrates the isolation of new <i>Acinetobacter</i>-infecting phages and the determination of their cell receptors. Further, we describe the genomes of a new genus of <i>Cystoviruses</i> and perform an initial characterization of membrane-associated glycoconjugates.https://www.mdpi.com/1999-4915/13/8/1652<i>Acinetobacter</i>bacteriophagessegmented RNA virusespilincapsular polysaccharideslipooligosaccharides |
| spellingShingle | Clay S. Crippen Bibi Zhou Silke Andresen Robert T. Patry Artur Muszyński Craig T. Parker Kerry K. Cooper Christine M. Szymanski RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 Viruses <i>Acinetobacter</i> bacteriophages segmented RNA viruses pilin capsular polysaccharides lipooligosaccharides |
| title | RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 |
| title_full | RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 |
| title_fullStr | RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 |
| title_full_unstemmed | RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 |
| title_short | RNA and Sugars, Unique Properties of Bacteriophages Infecting Multidrug Resistant <i>Acinetobacter radioresistens</i> Strain LH6 |
| title_sort | rna and sugars unique properties of bacteriophages infecting multidrug resistant i acinetobacter radioresistens i strain lh6 |
| topic | <i>Acinetobacter</i> bacteriophages segmented RNA viruses pilin capsular polysaccharides lipooligosaccharides |
| url | https://www.mdpi.com/1999-4915/13/8/1652 |
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