Characterization of Natronobacterium magadii phage ΦCh1, a unique archaeal phage containing DNA and RNA

A novel archaeal bacteriophage, ΦCh1, was isolated from a haloalkalophilic archaeon Natronobacterium magadii upon spontaneous lysis. The phage‐cured strain N. magadii (L13) was used to demonstrate infectivity of phage ΦCh1. The turbid‐plaque morphology and the fact that N. magadii cells isolated fro...

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Veröffentlicht in:Molecular microbiology 1997-02, Vol.23 (3), p.603-616
Hauptverfasser: Witte, Angela, Baranyi, Ulrike, Klein, Reinhard, Sulzner, Michael, Luo, Cheng, Wanner, Gerhard, Kru¨ger, Detlev H., Lubitz, Werner
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container_end_page 616
container_issue 3
container_start_page 603
container_title Molecular microbiology
container_volume 23
creator Witte, Angela
Baranyi, Ulrike
Klein, Reinhard
Sulzner, Michael
Luo, Cheng
Wanner, Gerhard
Kru¨ger, Detlev H.
Lubitz, Werner
description A novel archaeal bacteriophage, ΦCh1, was isolated from a haloalkalophilic archaeon Natronobacterium magadii upon spontaneous lysis. The phage‐cured strain N. magadii (L13) was used to demonstrate infectivity of phage ΦCh1. The turbid‐plaque morphology and the fact that N. magadii cells isolated from plaques were able to produce phage indicated that ΦCh1 is a temperate phage. The phage morphology resembles other members of Myoviridae‐infecting Halobacterium species. In solution below 2 M NaCl, the phage lost its morphological stability and infectivity. One‐ and two‐dimensional SDS–PAGE of phage particles revealed at least four major and five minor proteins with molecular masses ranging from 15 to 80 kDa and acidic isoelectric points. Southern blot analysis of chromosomal DNA of a lysogenic N. magadii strain showed that ΦCh1 exists as a chromosomally integrated prophage. The phage particles contain both double‐stranded, linear DNA (approx. 55 kbp) as well as several RNA species (80–700 nucleotides). Hybridization of labelled RNA fragments to total DNA from N. magadii and ΦCh1 showed that the virion‐associated RNA is host encoded. Part of the phage DNA population is modified and restriction analysis revealed evidence for adenine methylation. Phage ΦCh1 is the first virus described for the genus Natronobacterium, and the first phage containing DNA and RNA in mature phage particles.
doi_str_mv 10.1046/j.1365-2958.1997.d01-1879.x
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The phage‐cured strain N. magadii (L13) was used to demonstrate infectivity of phage ΦCh1. The turbid‐plaque morphology and the fact that N. magadii cells isolated from plaques were able to produce phage indicated that ΦCh1 is a temperate phage. The phage morphology resembles other members of Myoviridae‐infecting Halobacterium species. In solution below 2 M NaCl, the phage lost its morphological stability and infectivity. One‐ and two‐dimensional SDS–PAGE of phage particles revealed at least four major and five minor proteins with molecular masses ranging from 15 to 80 kDa and acidic isoelectric points. Southern blot analysis of chromosomal DNA of a lysogenic N. magadii strain showed that ΦCh1 exists as a chromosomally integrated prophage. The phage particles contain both double‐stranded, linear DNA (approx. 55 kbp) as well as several RNA species (80–700 nucleotides). Hybridization of labelled RNA fragments to total DNA from N. magadii and ΦCh1 showed that the virion‐associated RNA is host encoded. Part of the phage DNA population is modified and restriction analysis revealed evidence for adenine methylation. 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The phage‐cured strain N. magadii (L13) was used to demonstrate infectivity of phage ΦCh1. The turbid‐plaque morphology and the fact that N. magadii cells isolated from plaques were able to produce phage indicated that ΦCh1 is a temperate phage. The phage morphology resembles other members of Myoviridae‐infecting Halobacterium species. In solution below 2 M NaCl, the phage lost its morphological stability and infectivity. One‐ and two‐dimensional SDS–PAGE of phage particles revealed at least four major and five minor proteins with molecular masses ranging from 15 to 80 kDa and acidic isoelectric points. Southern blot analysis of chromosomal DNA of a lysogenic N. magadii strain showed that ΦCh1 exists as a chromosomally integrated prophage. The phage particles contain both double‐stranded, linear DNA (approx. 55 kbp) as well as several RNA species (80–700 nucleotides). Hybridization of labelled RNA fragments to total DNA from N. magadii and ΦCh1 showed that the virion‐associated RNA is host encoded. Part of the phage DNA population is modified and restriction analysis revealed evidence for adenine methylation. 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Hybridization of labelled RNA fragments to total DNA from N. magadii and ΦCh1 showed that the virion‐associated RNA is host encoded. Part of the phage DNA population is modified and restriction analysis revealed evidence for adenine methylation. Phage ΦCh1 is the first virus described for the genus Natronobacterium, and the first phage containing DNA and RNA in mature phage particles.</abstract><cop>Oxford BSL</cop><pub>Blackwell Science Ltd</pub><pmid>9044293</pmid><doi>10.1046/j.1365-2958.1997.d01-1879.x</doi><tpages>14</tpages></addata></record>
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subjects Archaea - virology
Bacteriophages - isolation & purification
Bacteriophages - physiology
Bacteriophages - ultrastructure
DNA, Viral - chemistry
DNA, Viral - physiology
DNA-Cytosine Methylases - metabolism
Electrophoresis, Agar Gel
Electrophoresis, Polyacrylamide Gel
Halobacterium
Natronobacterium
RNA, Viral - chemistry
RNA, Viral - physiology
Viral Proteins - chemistry
Viral Proteins - physiology
title Characterization of Natronobacterium magadii phage ΦCh1, a unique archaeal phage containing DNA and RNA
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