title: Exploring the mechanism of inhibition of human telomerase by cysteine-reactive compounds.
abstract: Telomerase is an almost universal cancer target that consists minimally of a core protein human telomerase reverse transcriptase (hTERT) and a RNA component human telomerase RNA (hTR). Some inhibitors of this enzyme are thought to function by the covalent binding to one or several cysteine residues; however, this inhibition mechanism has never been investigated because of the difficulty in producing telomerase. In this study, we use a recent method to produce recombinant hTERT to analyze the effect of cysteine-reactive inhibitors on telomerase. Using mass spectrometry and mutagenesis analysis, we identify several targeted residues in separated domains of the hTERT protein and show that cysteine-reactive reagents abolish the interaction with the CR4/5 region of hTR.
authors.affiliation: INSERM UMR-S 1007, Université Paris Descartes, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Kellermann
authors.affiliation: Laboratoire de Spectrometrie de Masse Protéomique, Institut Curie, University PSL, Paris, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Dingli
authors.affiliation: Laboratoire de Spectrometrie de Masse Protéomique, Institut Curie, University PSL, Paris, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Masson
authors.affiliation: Département Recherche, CNRS UMR3666/INSERM U1143, Institut Curie, Paris Cedex 05, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Dauzonne
authors.affiliation: INSERM UMR-S 1007, Université Paris Descartes, France.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Ségal-Bendirdjian
authors.affiliation: CNRS UMR9187/INSERM U1196, Institut Curie, Centre Universitaire, University Paris Sud, Orsay, France.
authors.forename: Marie-Paule
authors.initials: MP
authors.lastname: Teulade-Fichou
authors.affiliation: Laboratoire de Spectrometrie de Masse Protéomique, Institut Curie, University PSL, Paris, France.
authors.forename: Damarys
authors.initials: D
authors.lastname: Loew
authors.affiliation: INSERM UMR-S 1007, Université Paris Descartes, France.|CNRS UMR9187/INSERM U1196, Institut Curie, Centre Universitaire, University Paris Sud, Orsay, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bombard
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D003545:Cysteine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D015394:Molecular Structure; D016296:Mutagenesis; D011485:Protein Binding; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D019098:Telomerase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; C509186:TERT protein, human; D019098:Telomerase; D003545:Cysteine
keywords: cysteine; inhibitor; telomerase