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Alexandre Ceccaldi, Dominique Guianvarc'h, Catherine Senamaud-Beaufort, Renata Jurkowska, Daniel Dauzonne, Albert Jeltsch, Paola B Arimondo In quest of DNMT inhibitors not submitted contact: Dr Paola B Arimondo CNRS UMR5153 INSERM U565 MNHN USM503 arimondo@mnhn.fr 43 rue Cuvier 75005 Paris (France) additional information Alexandre Ceccaldi 1, Dominique Guianvarc'h 2, Catherine Senamaud-Beaufort 1, Renata Jurkowska 3, Daniel Dauzonne 4, Albert Jeltsch 3, Paola B. Arimondo 1 1UMR5153 CNRS-MNHN USM503; INSERM U565, 43 rue Cuvier 75005 Paris France; email: arimondo@mnhn.fr 2UMR7613 CNRS-UPMC, Université Pierre et Marie Curie, boîte 182, 4, place Jussieu, 75005 Paris, France 3Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany 4UMR 176 CNRS Institut Curie, 26 rue d'Ulm, 75005 Paris, France
Careen Katryniok, Bernd L. Sorg, Dieter Steinhilber INDUCTION OF HUMAN 5-LIPOXYGENASE GENE EXPRESSION BY THE HISTONE DEACETYLASE INHIBITOR TRICHOSTATIN A - INVESTIGATIONS ON THE MECHANISM 5-lipoxygenase (5-LO) is the key enzyme in the formation of leukotrienes which are important inflammatory mediators. The 5-LO promoter lacks a TATA or CCAT box, but posseses a unique GC-rich sequence (1). We have shown previously that epigenetic mechanisms play a role in the transcription of 5-lipoxygenase. The 5-LO transcription is silenced by DNA methylation (2,3) and activated by treatment with histone desacetylase inhibitior trichostatin A (TsA) (4). In this study, we continued the investigations concerning the activation of the 5-LO promoter by TsA. We explored the time dependency of the effect of TsA by rt-PCR and achieved the maximal effect 8-16 hours after incubation. Subsequently we analysed changes in the histone modifications of three different cell lines after TsA incubation, two of which are transcriptionally active (HL-60 and MM6 cells), whereas the third cell line is methylated and silenced (U937). We could discover differences between this cell lines in the acetylation pattern of the core histone proteins H3 and H4 using the chromatin immunoprecipitation assay. In TsA treated HL-60 and, surprisingly in U937 cells, we could detect an increase of acetylated Histon H3 and H4 with a maximum after 8-16 hours. Interestingly, no change in histone acetylation status is seen in the cell line MM6. It looks as if TsA does not act via the classical mechanism, which is increase of acetylation state of histone proteins, in MM6 cells. Literature 1. Hoshiko, S., Rådmark, O., and Samuelsson, B. (1990) Proceedings of the National Academy of Sciences of the United States of America 87, 9073-9077 2. Uhl, J., Klan, N., Rose, M., Entian, K. D., Werz, O., and Steinhilber, D. (2002) Journal of Biological Chemistry 277, 4374-4379 3. Uhl, J., Klan, N., Rose, M., Entian, K. D., Werz, O., and Steinhilber, D. (2003) Advances in Experimental Medicine & Biology 525, 169-172 4. Klan, N., Seuter, S., Schnur, N., Jung, M., and Steinhilber, D. (2003) Biological Chemistry 384, 777-785 contact: Careen Katryniok University of Frankfurt Institute of Pharmaceutical Chemistry Katryniok@pharmchem.uni-frankfurt.de Max-von-Laue-Str. 9 60438 Frankfurt (Germany)
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