distinct distribution of ectopically expressed histone variants h2a.bbd and macroh2a in open and closed chromatin domains不同分布的组蛋白变体h2a ectopically表达。.pdfVIP

Distinct Distribution of Ectopically Expressed Histone Variants H2A.Bbd and MacroH2A in Open and Closed Chromatin Domains Elena S. Ioudinkova1,2,3., Ana Barat2,4., Andrey Pichugin2,3, Elena Markova1,2,3, Ilya Sklyar1,2,3, Iryna Pirozhkova2,3, Chloe Robin2,3, Marc Lipinski2,3, Vasily Ogryzko2,3, Yegor S. Vassetzky2,3*, Sergey V. Razin1,3 ´ 1 Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia, 2 CNRS UMR 8126, Univ. Paris-Sud 11, Institut de cancerologie Gustave Roussy, Villejuif, France, 3 LIA1066, Laboratoire Franco-Russe de recherches en oncologie, Villejuif, France, 4 The Centre for Scientific Computing Complex Systems Modelling (SCI-SYM), School of Computing, Dublin City University, Dublin, Ireland Abstract Background: It becomes increasingly evident that nuclesomes are far from being identical to each other. This nucleosome diversity is due partially to the existence of histone variants encoded by separate genes. Among the known histone variants the less characterized are H2A.Bbd and different forms of macroH2A. This is especially true in the case of H2A.Bbd as there are still no commercially available antibodies specific to H2A.Bbd that can be used for chromatin immunoprecipitation (ChIP). Methods: We have generated HeLa S3 cell lines stably expressing epitope-tagged versions of macroH2A1.1, H2A.Bbd or canonical H2A and analyzed genomic distribution of the tagged histones using ChIP-on-chip technique. Results: The presence of histone H2A variants macroH2A1.1 and H2A.Bbd has been analyzed in the chromatin of several segments of human chromosomes 11, 16 and X that have been chosen for their different gene densities and chromatin status. Chromatin immunoprecipitation (ChIP) followed by hybridization with custom NimbleGe