The Chromatin Remodeling Protein, SRCAP, Is Critical for Deposition of the Histone Variant H2A.Z at Promoters

Department of Pharmacological and Physiological Science, Saint Louis University, Сент-Луис, Michigan, United States
Journal of Biological Chemistry (Impact Factor: 4.57). 10/2007; 282(36):26132-9. DOI: 10.1074/jbc.M703418200
Source: PubMed


Studies in Saccharomyces cerevisiae indicate the histone variant H2A.Z is deposited at promoters by the chromatin remodeling protein Swr1 and plays a critical role in the regulation of transcription. In higher eukaryotes, however, little is known about the distribution, method of deposition, and function of H2A.Z at promoters. Using biochemical studies, we demonstrated previously that SRCAP (SNF-2-related CREB-binding protein activator protein), the human ortholog of Swr1, could catalyze deposition of H2A.Z into nucleosomes. To address whether SRCAP directs H2A.Z deposition in vivo, promoters targeted by SRCAP were identified by a chromatin immunoprecipitation (ChIP)-on-chip assay. ChIP assays on a subset of these promoters confirmed the presence of SRCAP on inactive and active promoters. The highest levels of SRCAP were observed on the active SP-1, G3BP, and FAD synthetase promoters. Detailed analyses of these promoters indicate sites of SRCAP binding overlap or occur adjacent to the sites of H2A.Z deposition. Knockdown of SRCAP levels using siRNA resulted in loss of SRCAP at these promoters, decreased deposition of H2A.Z and acetylated H2A.Z, and a decrease in levels of SP-1, G3BP, and FAD synthetase mRNA. Thus, these studies provide the first evidence that SRCAP is recruited to promoters and is critical for the deposition of H2A.Z.

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    • "Different classes of genomic regions, such as promoters and enhancers, show discriminatory epigenetic patterns [25]. Regulatory mechanism operating at distinct classes often have a unique epigenetic component, such as the activity of a specific chromatin remodeling complex (e.g.[38]). Thus, it is reasonable to assume that specific or enriched combinatorial patterns could discriminate between classes of sites. "
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    ABSTRACT: Chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) is the most widely used method for characterizing the epigenetic states of chromatin on a genomic scale. With the recent availability of large genome-wide data sets, often comprising several epigenetic marks, novel approaches are required to explore functionally relevant interactions between histone modifications. Computational discovery of "chromatin states" defined by such combinatorial interactions enabled descriptive annotations of genomes, but more quantitative approaches are needed to progress towards predictive models. We propose non-negative matrix factorization (NMF) as a new unsupervised method to discover combinatorial patterns of epigenetic marks that frequently co-occur in subsets of genomic regions. We show that this small set of combinatorial "codes" can be effectively displayed and interpreted. NMF codes enable dimensionality reduction and have desirable statistical properties for regression and classification tasks. We demonstrate the utility of codes in the quantitative prediction of Pol2-binding and the discrimination between Pol2-bound promoters and enhancers. Finally, we show that specific codes can be linked to molecular pathways and targets of pluripotency genes during differentiation. We have introduced and evaluated a new computational approach to represent combinatorial patterns of epigenetic marks as quantitative variables suitable for predictive modeling and supervised machine learning. To foster widespread adoption of this method we make it available as an open-source software-package ¿ epicode at
    BMC Genomics 01/2014; 15(1):76. DOI:10.1186/1471-2164-15-76 · 3.99 Impact Factor
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    • "The ligand bound AR recruits co-activator proteins, including chromatin remodeling complexes that have histone posttranslational modification (PTM) activity, as well as components of the basic transcriptional machinery leading to gene expression. One such co-activator is SNF2-related CBP activator protein (SRCAP) that catalyzes the ATP-dependent incorporation of H2A.Z–H2B heterodimers into chromatin [5–7] and whose deregulation plays a critical role in prostate cancer [8]. "
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    ABSTRACT: Genetic and epigenetic changes are at the root of all cancers. The epigenetic component involves alterations of the post-synthetic modifications of DNA (methylation) and histones (histone posttranslational modifications, PTMs) as well as of those of their molecular "writers," "readers," and "erasers." Noncoding RNAs (ncRNA) can also play a role. Here, we focus on the involvement of histone alterations in cancer, in particular that of the histone variant H2A.Z in the etiology of prostate cancer. The structural mechanisms putatively responsible for the contribution of H2A.Z to oncogenic gene expression programs are first described, followed by what is currently known about the involvement of this histone variant in the regulation of androgen receptor regulated gene expression. The implications of this and their relevance to oncogene deregulation in different stages of prostate cancer, including the progression toward androgen independence, are discussed. This review underscores the increasing awareness of the epigenetic contribution of histone variants to oncogenic progression.
    CANCER AND METASTASIS REVIEW 01/2014; 33(2-3). DOI:10.1007/s10555-013-9486-9 · 7.23 Impact Factor
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    • "For example, review of the protein annotation (CCDS ID 10689) for the human SRCAP gene resulted in an N-terminal extension that adds an HSA domain that is found in DNA-binding proteins and is often associated with helicases. The HSA domain is consistent with the other domains found in the protein and with the presumed function of this protein as a component of the SRCAP chromatin remodeling complex ( Johnston et al. 1999; Wong et al. 2007). This significant improvement to the protein annotation was immediately available in the RefSeq transcript and protein sequences and in the Vega and UCSC Genome Browsers. "
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    ABSTRACT: Effective use of the human and mouse genomes requires reliable identification of genes and their products. Although multiple public resources provide annotation, different methods are used that can result in similar but not identical representation of genes, transcripts, and proteins. The collaborative consensus coding sequence (CCDS) project tracks identical protein annotations on the reference mouse and human genomes with a stable identifier (CCDS ID), and ensures that they are consistently represented on the NCBI, Ensembl, and UCSC Genome Browsers. Importantly, the project coordinates on manually reviewing inconsistent protein annotations between sites, as well as annotations for which new evidence suggests a revision is needed, to progressively converge on a complete protein-coding set for the human and mouse reference genomes, while maintaining a high standard of reliability and biological accuracy. To date, the project has identified 20,159 human and 17,707 mouse consensus coding regions from 17,052 human and 16,893 mouse genes. Three evaluation methods indicate that the entries in the CCDS set are highly likely to represent real proteins, more so than annotations from contributing groups not included in CCDS. The CCDS database thus centralizes the function of identifying well-supported, identically-annotated, protein-coding regions.
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