Identification and functional analysis of the Rz/Rz1-like accessory lysis genes in the membrane-containing bacteriophage PRD1.
Bacteriophage PRD1 is a tailless membrane-containing double-stranded (ds) DNA virus infecting a variety of Gram-negative bacteria. In order to affect cell lysis, like most dsDNA phages, PRD1 uses the holin-endolysin system. In this study, we identified two accessory lysis genes, XXXVI and XXXVII, coding for proteins P36 and P37, respectively. Using genetic complementation assays, we show that protein pair P36/P37 is a functional and interchangeable analogue of the Rz/Rz1 of bacteriophage lambda. Utilizing molecular biology, electrochemical as well as various microscopic techniques, we characterized the lysis phenotypes of PRD1 host cells infected with mutant viruses. Our results indicate that proteins P36 and P37 confer a competitive advantage to the phage by securing the efficient disruption of the infected cell and consequent release of the phage progeny under less favourable growth conditions. In concordance with prior data and the results obtained in this study, we propose a model explaining the role of Rz/Rz1-like proteins in the lysis process: Rz/Rz1 complexes transform the mechanical stress caused by the holin lesion at the CM to the OM leading to its disintegration. Finally, identification of the Rz/Rz1-like genes in PRD1 suggests that tailless icosahedral phages are involved in genetic trade with tailed bacteriophages.
Krupovic
Mart
M
Department of Biological and Environmental Sciences and Institute of Biotechnology, Biocenter 2, PO Box 56 (Viikinkaari 5), FIN-00014, University of Helsinki, Helsinki, Finland.
Cvirkaite-Krupovic
Virginija
V
Bamford
Dennis H
DH
D000595:Amino Acid Sequence; D001433:Bacteriolysis; D025622:Bacteriophage PRD1 / Q000201:enzymology* / Q000235:genetics; D010582:Bacteriophage lambda / Q000201:enzymology / Q000235:genetics; D003067:Coenzymes / Q000494:pharmacology; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D048168:Escherichia coli K12 / Q000821:virology; D023061:Gene Order; D005816:Genetic Complementation Test; D015636:Magnesium Chloride / Q000494:pharmacology; D008954:Models, Biological; D008969:Molecular Sequence Data; D012486:Salmonella typhimurium / Q000166:cytology / Q000821:virology; D016415:Sequence Alignment; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D003067:Coenzymes; D014764:Viral Proteins; D015636:Magnesium Chloride; D010450:Endopeptidases; C001143:endolysin