Photolyase-dimer-DNA complexes and exclusion stimulation in Escherichia coli: depolarization of the plasma membrane.
Using cells that overproduce DNA photolyase, we found that UV irradiation (3 J/m2) efficiently inactivates accumulation of methylthiogalactoside (TMG) when RexAB proteins of phage lambda are present. The effect requires both formation of photolyase-dimer-DNA (PDD) complexes and expression of the RexAB proteins. It is reversed completely by a flash of visible light if given immediately after UV and becomes irreversible after post-UV incubation for about 15 min. Inactivation is significant after only 5 min of post-UV incubation, is accompanied by a loss of previously accumulated TMG, and does not require de novo protein synthesis. Passive transport of O-nitrophenylgalactoside by inactivated cells is typical of energy-depleted membranes. We suggest that PDD complexes mimic a developmental intermediate of phage superinfection and stimulate formation of the RexB membrane channel recently proposed by others to explain classical "exclusion". This suggestion is supported by additional data showing an inactivation of colony-forming ability by exclusion stimulation and an inability of PDD complexes to inactivate accumulation of TMG if RexB is present in larger relative amounts than RexA (a detail characteristic of exclusion stimulated by phage superinfection).
Li
B H
BH
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202.
Bockrath
R
R
D010582:Bacteriophage lambda / Q000378:metabolism; D002462:Cell Membrane / Q000502:physiology; D004269:DNA, Bacterial / Q000378:metabolism*; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism* / Q000528:radiation effects; D008564:Membrane Potentials; D008756:Methylgalactosides / Q000378:metabolism; D013862:Thiogalactosides / Q000378:metabolism; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D004269:DNA, Bacterial; D008756:Methylgalactosides; D013862:Thiogalactosides; D014764:Viral Proteins; C069331:rexB protein, Bacteriophage lambda; C009468:thiomethylgalactoside; D004255:Deoxyribodipyrimidine Photo-Lyase