Pentamidine-induced alteration in restriction endonuclease cleavage of plasmid DNA.
We have used restriction enzymes and DNaseI as probes to determine the specificity of pentamidine binding to plasmid DNA. Cleavage of plasmid pAZ130 by EcoRI, EcoRV and ApaI is inhibited by pentamidine, cleavage by XbaI, NotI and AvaI is unaffected, while cleavage by XhoI, which recognizes the same sequence as AvaI, is stimulated. DNaseI footprinting of DNA containing these restriction sites revealed that pentamidine protection is not strictly limited to AT-rich regions. We suggest that perturbation of the DNA micro- environment by pentamidine binding is responsible for its effect on nucleases.
Choudhury
Kanakendu
K
Department of Molecular Genetics, Microbiology and Immunology, Robert Wood Johnson Medical School-UMDNJ, Piscataway, NJ 08854, USA.
Leibowitz
Michael J
MJ
D001482:Base Composition / Q000187:drug effects / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D004262:DNA Restriction Enzymes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D005822:Genetic Vectors; D007364:Intercalating Agents / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D010419:Pentamidine / Q000378:metabolism / Q000494:pharmacology*; D010957:Plasmids / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
D004269:DNA, Bacterial; D007364:Intercalating Agents; D010419:Pentamidine; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I