tailieunhanh - Báo cáo y học: "Quantitative analysis of histone exchange for transcriptionally active chromatin"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Wertheim cung cấp cho các bạn kiến thức về ngành y đề tài: Quantitative analysis of histone exchange for transcriptionally active chromatin. | Byrum et al. Journal of Clinical Bioinformatics 2011 1 17 http content 1 1 17 JOURNAL OF CLINICAL BIOINFORMATICS SHORT REPORT Open Access Quantitative analysis of histone exchange for transcriptionally active chromatin Stephanie D Byrum1 Sean D Taverna2t and Alan J Tackett1 Abstract Background Genome-wide studies use techniques like chromatin immunoprecipitation to purify small chromatin sections so that protein-protein and protein-DNA interactions can be analyzed for their roles in modulating gene transcription. Histone post-translational modifications PTMs are key regulators of gene transcription and are therefore prime targets for these types of studies. Chromatin purification protocols vary in the amount of chemical cross-linking used to preserve in vivo interactions. A balanced level of chemical cross-linking is required to preserve the native chromatin state during purification while still allowing for solubility and interaction with affinity reagents. Findings We previously used an isotopic labeling technique combining affinity purification and mass spectrometry called transient isotopic differentiation of interactions as random or targeted transient I-DIRT to identify the amounts of chemical cross-linking required to prevent histone exchange during chromatin purification. New bioinformatic analyses reported here reveal that histones containing transcription activating PTMs exchange more rapidly relative to bulk histones and therefore require a higher level of cross-linking to preserve the in vivo chromatin structure. Conclusions The bioinformatic approach described here is widely applicable to other studies requiring the analysis and purification of cognate histones and their modifications. Histones containing PTMs correlated to active gene transcription exchange more readily than bulk histones therefore it is necessary to use more rigorous in vivo chemical cross-linking to stabilize these marks during chromatin purification. .

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