Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer: inhibition of the elongation of a telomeric repeat by telomerase.
The binding of a dimeric form of the 2-amino-1,8-naphthyridine derivative (naphthyridine dimer) to a human telomeric sequence, TTAGGG, was investigated by UV melting, CD spectra, and CSI-MS measurements. Both the 9-mer d(TTAGGGTTA) and the 15-mer d(TTAGGGTTAGGGTTA) showed apparent melting temperatures (T(m)) of 45.6 and 63.6 degrees C, respectively, in the presence of naphthyridine dimer (30 microM) in sodium cacodylate buffer (50 mM, pH 7.0) containing 100 mM NaCl. The CD spectra at 235 and 255 nm of the 9-mer increased in intensity accompanied with strong induced CDs at 285 and 340 nm upon complex formation with naphthyridine dimer. UV titration of the binding of naphthyridine dimer to the 9-mer at 320 nm showed a hypochromism of the spectra. A Scatchard plot of the data showed the presence of multiple binding sites with different association constants. Cold spray ionization mass spectrometry of the complex between naphthyridine dimer and the 9-mer clearly showed that one to three molecules of the ligand bound to the dimer duplex of the 9-mer. Telomeric repeat elongation assay showed that the binding of naphthyridine dimer to the telomeric sequence inhibits the elongation of the sequence by telomerase.
Nakatani
Kazuhiko
K
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Japan. nakatani@sbchem.kyoto-u.ac.jp
Hagihara
Shinya
S
Sando
Shinsuke
S
Sakamoto
Shigeru
S
Yamaguchi
Kentaro
K
Maesawa
Chihaya
C
Saito
Isao
I
D020137:Base Pair Mismatch; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009287:Naphthyridines / Q000737:chemistry* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D012091:Repetitive Sequences, Nucleic Acid; D019098:Telomerase / Q000737:chemistry* / Q000378:metabolism; D016615:Telomere / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D009287:Naphthyridines; D004247:DNA; D019098:Telomerase