TY - JOUR
T1 - Rna and sugars, unique properties of bacteriophages infecting multidrug resistant acinetobacter radioresistens strain lh6
AU - Crippen, Clay S.
AU - Zhou, Bibi
AU - Andresen, Silke
AU - Patry, Robert T.
AU - Muszyński, Artur
AU - Parker, Craig T.
AU - Cooper, Kerry K.
AU - Szymanski, Christine M.
N1 - Funding Information:
C.T.P. was supported by the United States Department of Agriculture, Agricultural Research Service CRIS project 2030-42000-055-000-D from the (https://www.ars.usda.gov/research/ project/?accnNo=440168, accessed on 15 April 2021). Work by AM at the Complex Carbohydrate Research Center was supported in part by NIH grant R24GM137782-01 to Parastoo Azadi.
Funding Information:
Funding: C.T.P. was supported by the United States Department of Agriculture, Agricultural Research Service CRIS project 2030-42000-055-000-D from the (https://www.ars.usda.gov/research/ project/?accnNo=440168, accessed on 15 April 2021). Work by AM at the Complex Carbohydrate Research Center was supported in part by NIH grant R24GM137782-01 to Parastoo Azadi.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/8
Y1 - 2021/8
N2 - 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 Acinetobacter-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 Acinetobacter radioresistens 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 Pseudomonas species. In Pseudomonas, 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 Acinetobacter-infecting phages and the determination of their cell receptors. Further, we describe the genomes of a new genus of Cystoviruses and perform an initial characterization of membrane-associated glycoconjugates.
AB - 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 Acinetobacter-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 Acinetobacter radioresistens 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 Pseudomonas species. In Pseudomonas, 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 Acinetobacter-infecting phages and the determination of their cell receptors. Further, we describe the genomes of a new genus of Cystoviruses and perform an initial characterization of membrane-associated glycoconjugates.
KW - Acinetobacter
KW - Bacteriophages
KW - Capsular polysaccha-rides
KW - Lipooligosaccharides
KW - Pilin
KW - Segmented RNA viruses
UR - http://www.scopus.com/inward/record.url?scp=85113578875&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85113578875&partnerID=8YFLogxK
U2 - 10.3390/v13081652
DO - 10.3390/v13081652
M3 - Article
C2 - 34452516
AN - SCOPUS:85113578875
SN - 1999-4915
VL - 13
JO - Viruses
JF - Viruses
IS - 8
M1 - 1652
ER -