BIOL 7 |
| A complex interplay between DNA, histones, and chromatin associated proteins coordinates the structure and function of the eukaryotic genome. To help accomplish this task, extensive posttranslational modifications of the core histone proteins have evolved. One of the less elucidated modifications, monoubiquitylation of H2B (uH2B), has been implicated in transcription elongation and histone methylation of H3 K4 and K79. Characterization of the role of uH2B in these processes requires the isolation or generation of homogenously ubiquitylated H2B. To this end, we have developed a traceless semisynthetic strategy to generate uH2B. We employ two expressed protein ligation (EPL) reactions and two methods for ligation site removal to synthesize native uH2B. The resulting uH2B was reconstituted into core histone octamers and nucleosome arrays were then generated for biochemical characterization. Synthetic and biochemical aspects of this work will be discussed. |
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Chemical Approaches to Protein Function
2:00 PM-4:40 PM, Sunday, August 19, 2007 BCEC -- 109A, Oral
Division of Biological Chemistry |