부산대학교 도서관

To purchase or authenticate to the full-text of this article, please visit this link: http://dx.doi.org/10.1111/j.1365-2958.2007.06079.x Byline: T. Nicolai Siegel (1), Taemi Kawahara (2), Jeffrey A. DeGrasse (3), Christian J. Janzen (1[dagger]), David Horn (2), George A. M. Cross (1) Abstract: Summary Post-translational histone modifications have been studied intensively in several eukaryotes. It has been proposed that these modifications constitute a 'histone code' that specifies epigenetic information for transcription regulation. With a limited number of histone-modifying enzymes, implying less redundancy, Trypanosoma brucei represents an excellent system in which to investigate the function of individual histone modifications and histone-modifying enzymes. In this study, we characterized the acetylation of lysine 4 of histone H4 (H4K4), the most abundant acetylation site in T. brucei histones. Because of the large sequence divergence of T. brucei histones, we generated highly specific antibodies to acetylated and unmodified H4K4. Immunofluorescence microscopy and Western blots with sorted cells revealed a strong enrichment of unmodified H4K4 in S phase and suggested a G1/G0-specific masking of the site, owing to non-covalently binding factors. Finally, we showed that histone acetyltransferase 3 (HAT3) is responsible for H4K4 acetylation and that treatment of cells with the protein synthesis inhibitor cycloheximide led to an almost instantaneous loss of unmodified H4K4 sites. As HAT3 is located inside the nucleus, our findings suggest that newly synthesized histone H4 with an unmodified K4 is imported rapidly into the nucleus, where it is acetylated, possibly irreversibly. Author Affiliation: (1)Laboratory of Molecular Parasitology, The Rockefeller University, New York, USA. (2)London School of Hygiene and Tropical Medicine, London, UK. (3)Laboratory for Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, USA. Article History: Accepted 6 December, 2007. Article note: (*) E-mail gamc@rockefeller.edu; Tel. (+1) 212 327 7571; Fax (+1) 212 327 7845.