Phage HK022 Roi protein inhibits phage lytic growth in Escherichia coli integration host factor mutants.
Temperate coliphage HK022 requires integration host factor (IHF) for lytic growth. The determinant responsible for this requirement was identified as a new gene (roi) located between genes P and Q. This gene encodes a DNA-binding protein (Roi) containing a helix-turn-helix motif. We have shown that Roi binds a site within its own gene that is closely linked to an IHF binding site. By gel retardation experiments, we have found that IHF binding stabilizes the interaction of Roi with its gene. We have isolated three independent phage mutants that are able to grow on an IHF- host. They carry different mutations scattered in the roi gene and specifying single amino-acid changes. The interactions of all three Roi mutant proteins with the Roi binding site differed from that of the wild type. Roi displays strong similarities, in its C-terminal half, to two putative DNA-binding proteins of bacteriophage P1: Ant1 and KilA. The mode of action of the Roi protein and the possibility that IHF is modulating the expression and/or the action of Roi are discussed.
Clerget
M
M
Département de Biologie Moléculaire, Université de Genève, Switzerland.
Boccard
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F
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D010582:Bacteriophage lambda / Q000254:growth & development*; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D005814:Genes, Viral; D005819:Genetic Markers; D034982:Integration Host Factors; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000378:metabolism*
D001426:Bacterial Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D034982:Integration Host Factors; C100964:Roi protein, Bacteriophage HK022; D014764:Viral Proteins