In vitro DNA crosslinking by Ledakrin, an antitumor derivative of 1-nitro-9-aminoacridine.
Using agarose gel electrophoresis we confirmed that Ledakrin is capable of incurring covalent crosslinking in pBR322 plasmid DNA and also in poly(dGdC) in the presence of a simple activating system containing DTT. The identification of adducts resulting from DNA crosslinking was carried out by 32P-post-labelling assay. We assumed that such adduct(s) should be brought about more readily with double-stranded than with single-stranded polynucleotides or nucleotides. Since our earlier experiments had shown that guanine is a major site of covalent binding of 1-nitroacridines, we compared DNA adduct formation by Ledakrin for ctDNA, dG-containing synthetic homopolymers and 3'-pdG. 32P-Post-labelling assay revealed two adduct spots that were enhanced in samples containing double-stranded substrates in which interstrand crosslinking between guanines was possible, namely ctDNA and poly(dGdC).
Bartoszek
A
A
Department of Pharmaceutical Technology and Biochemistry, Technical University of Gdańsk, Poland.
Dackiewicz
P
P
Składanowski
A
A
Konopa
J
J
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D003432:Cross-Linking Reagents / Q000378:metabolism / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects* / Q000378:metabolism*; D018736:DNA Adducts / Q000096:biosynthesis*; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D007364:Intercalating Agents / Q000378:metabolism / Q000494:pharmacology*; D009564:Nitracrine / Q000378:metabolism / Q000494:pharmacology*; D011066:Poly C / Q000378:metabolism; D011068:Poly G / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000378:metabolism
D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D018736:DNA Adducts; D004277:DNA, Single-Stranded; D007364:Intercalating Agents; D013439:Sulfhydryl Reagents; D011068:Poly G; D011066:Poly C; D009564:Nitracrine; D004247:DNA; D004229:Dithiothreitol