000176173 001__ 176173
000176173 005__ 20181203022657.0
000176173 0247_ $$2doi$$a10.1016/j.biochi.2011.12.004
000176173 02470 $$2ISI$$a000301332200032
000176173 037__ $$aARTICLE
000176173 245__ $$aSpecific binding of telomeric G-quadruplexes by hydrosoluble perylene derivatives inhibits repeat addition processivity of human telomerase
000176173 269__ $$a2012
000176173 260__ $$c2012
000176173 336__ $$aJournal Articles
000176173 520__ $$aTelomerase is responsible for the immortal phenotype of cancer cells and telomerase inhibition may specifically target cancer cell proliferation. Ligands able to selectively bind to G-quadruplex telomeric DNA have been considered as telomerase inhibitors but their mechanisms of action have often been deduced from a non-quantitative telomerase activity assay (TRAP assay) that involves a PCR step and that does not provide insight on the mechanism of inhibition. Furthermore, quadruplex ligands have also been shown to exert their effects by affecting association of telomere binding proteins with telomeres. Here, we use quantitative direct telomerase activity assays to evaluate the strength and mechanism of action of hydrosoluble perylene diimides (HPDIs). HPDIs contain a perylene moiety and different numbers of positively charged side chains. Side chain features vary with regard to number and distances of the charges. IC50 values of HPDIs were in the low micromolar (0.5-5 mu M) range depending on the number and features of the side chains. HPDIs having four side chains emerged as the best compounds of this series. Analysis of primer elongation products demonstrated that at low HPDI concentrations, telomerase inhibition involved formation of telomeric G-quadruplex structures, which inhibited further elongation by telomerase. At high HPDI concentrations, telomerase inhibition occurred independently of G-quadruplex formation of the substrate. The mechanism of action of HPDIs and their specific binding to G-quadruplex DNA was supported by PAGE analysis, CD spectroscopy and ESI-MS. Finally, competition Telospot experiments with duplex DNA indicated specific binding of HPDIs to the single-stranded telomeric substrates over double stranded DNA, a result supported by competitive ESI-MS. Altogether, our results indicate that HPDIs act by stabilizing G-quadruplex structures in single-stranded telomeric DNA, which in turn prevents repeat addition processivity of telomerase. (C) 2011 Elsevier Masson SAS. All rights reserved.
000176173 6531_ $$aG-quadruplex
000176173 6531_ $$aPerylene diimide
000176173 6531_ $$aTelomerase
000176173 6531_ $$aTelomere
000176173 6531_ $$aInhibitor
000176173 6531_ $$aTelospot
000176173 6531_ $$aDifferent Side-Chains
000176173 6531_ $$aDna Structures
000176173 6531_ $$aReverse-Transcriptase
000176173 6531_ $$aCoronene Derivatives
000176173 6531_ $$aMass-Spectrometry
000176173 6531_ $$aLigands
000176173 6531_ $$aCancer
000176173 6531_ $$aCells
000176173 6531_ $$aPolymorphism
000176173 6531_ $$aCompound
000176173 700__ $$aD'Ambrosio, Danilo
000176173 700__ $$aReichenbach, Patrick
000176173 700__ $$aMicheli, Emanuela
000176173 700__ $$aAlvino, Antonello
000176173 700__ $$aFranceschin, Marco
000176173 700__ $$aSavino, Maria
000176173 700__ $$g168670$$aLingner, Joachim$$0240570
000176173 773__ $$j94$$tBiochimie$$q854-863
000176173 909C0 $$xU11159$$0252147$$pUPLIN
000176173 909CO $$pSV$$particle$$ooai:infoscience.tind.io:176173
000176173 917Z8 $$x168670
000176173 937__ $$aEPFL-ARTICLE-176173
000176173 973__ $$rREVIEWED$$sPUBLISHED$$aEPFL
000176173 980__ $$aARTICLE