title: Identification of human telomerase assembly inhibitors enabled by a novel method to produce hTERT.
abstract: Telomerase is the enzyme that maintains the length of telomeres. It is minimally constituted of two components: a core reverse transcriptase protein (hTERT) and an RNA (hTR). Despite its significance as an almost universal cancer target, the understanding of the structure of telomerase and the optimization of specific inhibitors have been hampered by the limited amount of enzyme available. Here, we present a breakthrough method to produce unprecedented amounts of recombinant hTERT and to reconstitute human telomerase with purified components. This system provides a decisive tool to identify regulators of the assembly of this ribonucleoprotein complex. It also enables the large-scale screening of small-molecules capable to interfere with telomerase assembly. Indeed, it has allowed us to identify a compound that inhibits telomerase activity when added prior to the assembly of the enzyme, while it has no effect on an already assembled telomerase. Therefore, the novel system presented here may accelerate the understanding of human telomerase assembly and facilitate the discovery of potent and mechanistically unique inhibitors.
authors.affiliation: INSERM UMR-S 1007, Cellular Homeostasis and Cancer, Paris, France Université Paris Descartes, Paris Sorbonne Cité, Paris, France guikell@gmail.com.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Kellermann
authors.affiliation: Institut Curie, CMIB, CNRS UMR 9187- INSERM U1196, Orsay, France.
authors.forename: Markus
authors.initials: M
authors.lastname: Kaiser
authors.affiliation: Institut Curie/laboratoire de spectrométrie de masse protéomique, Paris, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Dingli
authors.affiliation: INSERM U-1016, Institut Cochin, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Lahuna
authors.affiliation: Institut Curie, CMIB, CNRS UMR 9187- INSERM U1196, Orsay, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Naud-Martin
authors.affiliation: Institut Curie, CMIB, CNRS UMR 9187- INSERM U1196, Orsay, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Mahuteau-Betzer
authors.affiliation: Institut Curie/laboratoire de spectrométrie de masse protéomique, Paris, France.
authors.forename: Damarys
authors.initials: D
authors.lastname: Loew
authors.affiliation: INSERM UMR-S 1007, Cellular Homeostasis and Cancer, Paris, France Université Paris Descartes, Paris Sorbonne Cité, Paris, France.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Ségal-Bendirdjian
authors.affiliation: Institut Curie, CMIB, CNRS UMR 9187- INSERM U1196, Orsay, France.
authors.forename: Marie-Paule
authors.initials: MP
authors.lastname: Teulade-Fichou
authors.affiliation: INSERM UMR-S 1007, Cellular Homeostasis and Cancer, Paris, France Université Paris Descartes, Paris Sorbonne Cité, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bombard
mesh_terms: D000166:Acridines / Q000737:chemistry / Q000494:pharmacology; D005821:Genetic Techniques; D057809:HEK293 Cells; D006801:Humans; D009536:Niacinamide / Q000031:analogs & derivatives; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D059370:RNA Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship; D019098:Telomerase / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles; D014312:Trisaccharides / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000166:Acridines; D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; C000591009:TAI-1 compound; D013844:Thiazoles; D014312:Trisaccharides; C422952:telomerase RNA; D009536:Niacinamide; D012313:RNA; C509186:TERT protein, human; D019098:Telomerase