Bacteriophage P22 Abc2 protein binds to RecC increases the 5' strand nicking activity of RecBCD and together with lambda bet, promotes Chi-independent recombination.
Bacteriophage P22 Abc2 protein binds to the RecBCD enzyme from Escherichia coli to promote phage growth and recombination. Overproduction of the RecC subunit in vivo, but not RecB or RecD, interfered with Abc2-induced UV sensitization, revealing that RecC is the target for Abc2 in vivo. UV-induced ATP crosslinking experiments revealed that Abc2 protein does not interfere with the binding of ATP to either the RecB or RecD subunits in the absence of DNA, though it partially inhibits RecBCD ATPase activity. Productive growth of phage P22 in wild-type Salmonella typhimurium correlates with the presence of Abc2, but is independent of the absolute level of ATP-dependent nuclease activity, suggesting a qualitative change in the nature of Abc2-modified RecBCD nuclease activity relative to the native enzyme. In lambda phage crosses, Abc2-modified RecBCD could substitute for lambda exonuclease in Red-promoted recombination; lambda Gam could not. In exonuclease assays designed to examine the polarity of digestion, Abc2 protein qualitatively changes the nature of RecBCD double-stranded DNA exonuclease by increasing the rate of digestion of the 5' strand. In this respect, Abc2-modified RecBCD resembles a RecBCD molecule that has encountered the recombination hotspot Chi. However, unlike Chi-modified RecBCD, Abc2-modified RecBCD still possesses 3' exonuclease activity. These results are discussed in terms of a model in which Abc2 converts the RecBCD exonuclease for use in the P22 phage recombination pathway. This mechanism of P22-mediated recombination distinguishes it from phage lambda recombination, in which the phage recombination system (Red) and its anti-RecBCD function (Gam) work independently.
Murphy
K C
KC
Department of Molecular Genetics, University of Massachusetts Medical School, Worcester, MA, 01655, USA.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D017100:Bacteriophage P22 / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D010582:Bacteriophage lambda / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D003433:Crosses, Genetic; D004249:DNA Damage / Q000235:genetics / Q000528:radiation effects; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000821:virology; D029968:Escherichia coli Proteins; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D007700:Kinetics; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011836:Radiation Tolerance; D011995:Recombination, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics / Q000502:physiology; D012486:Salmonella typhimurium / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000821:virology; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D002352:Carrier Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014764:Viral Proteins; C405525:abc2 protein, bacteriophage P22; C118730:beta protein, Bacteriophage lambda; D000255:Adenosine Triphosphate; D005090:Exodeoxyribonucleases; C043586:exo protein, Bacteriophage lambda; D043211:Exodeoxyribonuclease V; C468322:exodeoxyribonuclease V, E coli; D000251:Adenosine Triphosphatases