Probing the surface of Z-DNA with anti-nucleoside antibodies.
Antibodies specific for cytidine (C) and guanosine (G) were used to probe the surface of two Z-DNA conformers. When tested by ELISA, anti-G reacted with poly(dG-dC).poly(dG-dC) treated with bromine water [Br-poly(dG-dC).poly(dG-dC)] but anti-C did not. A weak reaction with anti-C was detected by dot immunobinding. In contrast, anti-C reacted strongly with poly(dG-dC).poly(dG-dC) treated with N-acetoxy-2-(acetylamino)fluorene [AAF-poly(dG-dC).poly(dG-dC)]; anti-G reacted weakly, despite the fact that most G residues had not been substituted with AAF. Neither antinucleoside bound to the B conformation of poly(dG-dC).poly(dG-dC). In competition experiments, GMP was the most efficient competitor of the reaction of anti-G with Br-poly(dG-dC).poly(dG-dC); AMP and TMP were 100-fold less efficient, and CMP did not compete to a significant extent. In contrast, the reaction of anti-Z with Br-poly(dG-dC).poly(dG-dC) was not inhibited by nucleotides. Of five possible sites recognized on guanosine by anti-G antibodies (N1, C6, O6, N7, and C8), AMP and TMP share three or their equivalent and CMP only one. The binding of anti-C to AAF-poly(dG-dC).poly(dG-dC) was inhibited best by CMP; AMP was 8 times less efficient; GMP and TMP were about 35-fold less efficient than CMP. Thus, although the amino group on the C4 position of CMP appears to be immunodominant, the capacity of GMP and TMP to inhibit the reaction indicates that other sites are also recognized in AAF-poly(dG-dC).poly(dG-dC), e.g., the exposed C5 position.(ABSTRACT TRUNCATED AT 250 WORDS)
Nahon
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E
Department of Microbiology, Columbia University, New York, New York 10032.
Grunberger
D
D
Erlanger
B F
BF
D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D001966:Bromine; D003562:Cytidine / Q000276:immunology*; D003568:Cytidine Monophosphate / Q000378:metabolism; D004247:DNA / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006151:Guanosine / Q000276:immunology*; D006157:Guanosine Monophosphate / Q000378:metabolism; D006801:Humans; D009690:Nucleic Acid Conformation; D011089:Polydeoxyribonucleotides / Q000378:metabolism; D011817:Rabbits; D013499:Surface Properties; D013938:Thymidine Monophosphate / Q000378:metabolism
D011089:Polydeoxyribonucleotides; D006151:Guanosine; C019530:poly(dC-dG); D013938:Thymidine Monophosphate; D000249:Adenosine Monophosphate; D003562:Cytidine; D006157:Guanosine Monophosphate; D004247:DNA; D003568:Cytidine Monophosphate; D001966:Bromine