The Histone Mark H3K36me3 Regulates Human DNA Mismatch Repair through Its Interaction with MutSalpha

Graduate Center for Toxicology, Markey Cancer Center, University of Kentucky College of Medicine, Lexington, KY 40506, USA.
Cell (Impact Factor: 32.24). 04/2013; 153(3):590-600. DOI: 10.1016/j.cell.2013.03.025
Source: PubMed


DNA mismatch repair (MMR) ensures replication fidelity by correcting mismatches generated during DNA replication. Although human MMR has been reconstituted in vitro, how MMR occurs in vivo is unknown. Here, we show that an epigenetic histone mark, H3K36me3, is required in vivo to recruit the mismatch recognition protein hMutSα (hMSH2-hMSH6) onto chromatin through direct interactions with the hMSH6 PWWP domain. The abundance of H3K36me3 in G1 and early S phases ensures that hMutSα is enriched on chromatin before mispairs are introduced during DNA replication. Cells lacking the H3K36 trimethyltransferase SETD2 display microsatellite instability (MSI) and an elevated spontaneous mutation frequency, characteristic of MMR-deficient cells. This work reveals that a histone mark regulates MMR in human cells and explains the long-standing puzzle of MSI-positive cancer cells that lack detectable mutations in known MMR genes.

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    • "Recently, Boros et al. found that H3K9me plays a pivotal role in the formation of the heterochromatin and also contributes to the maintenance of gene silencing. Alternatively, it has been demonstrated that H3K36me has an ability to modulate genomic stability and cancer susceptibility through the regulation of DNA mismatch repair (Li et al. 2013). Gene amplified in squamous cell carcinoma (SCC) 1 (GASC1), which belongs to the JMJD2 subfamily, encodes a histone demethylase containing two PHD-finger motifs and a PX domain and catalyzes the demethylation of H3K9me and H3K36me (Yang et al. 2001) as well as of H1.4K26 (Trojer et al. 2009). "
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    ABSTRACT: Gene amplified in squamous cell carcinoma (SCC) 1 (GASC1), also known as KDM4C/JMJD2C, encodes a histone demethylase that specifically demethylates lysine residues (H3K9, H3K36, and H1.4K26) and plays a crucial role in the regulation of gene expression as well as in heterochromatin formation. GASC1 is located at human chromosome 9p23-24, where frequent genomic amplification is observed in human esophageal cancer, and its aberrant expression is detected in a variety of human cancers, such as breast, colon, and prostate. Therefore, it is highly likely that GASC1 contributes to the genesis and/or development of cancer. However, there is a lack of direct evidence of GASC1 having an oncogenic function. In this study, we aimed to clarify the role of GASC1 in the skin SCC carcinogenesis. For this purpose, we generated Gasc1-heterozygous mice (Gasc1 (+/-)) with reduced expression of Gasc1. On the basis of our results, Gasc1 (+/-) mice displayed a significantly lower incidence and multiplicity of both benign and malignant tumors induced by the two-stage skin carcinogenesis protocol than wild-type mice. In addition, the volume of carcinoma was significantly lower in Gasc1 (+/-) mice. Consistent with these observations, knocking down of Gasc1 resulted in reduced cell viability of SCC cells in vitro. Our findings clearly demonstrated that GASC1 has an oncogenic role in skin carcinogenesis.
    Mammalian Genome 08/2015; DOI:10.1007/s00335-015-9592-9 · 3.07 Impact Factor
    • "CGIs are generally unmethylated, and are located at 72% of annotated promoters in the human genome (Saxonov et al. 2006). Interestingly, regions of low CpG methylation can also be found for distal CGIs in enhancers (Stadler et al. 2011; Lister et al. 2009; Ziller et al. 2013). "
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    ABSTRACT: The intestinal epithelium is an ideal model system for the study of normal and pathological differentiation processes. The mammalian intestinal epithelium is a single cell layer comprising proliferative crypts and differentiated villi. The crypts contain both proliferating and quiescent stem cell populations that self-renew and produce all the differentiated cell types, which are replaced every 3-5 days. The genetics of intestinal development, homeostasis, and disease are well defined, but less is known about the contribution of epigenetics in modulating these processes. Epigenetics refers to heritable phenotypic traits, including gene expression, which are independent of mutations in the DNA sequence. We have known for several decades that human colorectal cancers contain hypomethylated DNA, but the causes and consequences of this phenomenon are not fully understood. In contrast, tumor suppressor gene promoters are often hypermethylated in colorectal cancer, resulting in decreased expression of the associated gene. In this review, we describe the role that epigenetics plays in intestinal homeostasis and disease, with an emphasis on results from mouse models. We highlight the importance of producing and analyzing next-generation sequencing data detailing the epigenome from intestinal stem cell to differentiated intestinal villus cell.
    Cellular and Molecular Life Sciences CMLS 07/2015; 72(21). DOI:10.1007/s00018-015-1997-9 · 5.81 Impact Factor
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    • "It is shown that Dnmt3A specifically recognizes and interacts with methylated H3K36 through its PWWP domain, and this interaction is necessary for proper DNA methylation [26]. In mammalians, it has been shown that trimethylated H3K36 is involved in DNA mismatch repair process through recruiting the hMutSα to the chromatin through interaction with PWWP domain of hMSH6 [8]. Although the methyltransferase activities of ASH1L and SETD2 have been previously reported [11] [21] [27] [28] [29], they haven't been fully characterized in vitro and their kinetic parameters have not been determined . "
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    ABSTRACT: Dysregulation of methylation of lysine 36 on histone H3 (H3K36) has been implicated in a variety of diseases including cancers. ASH1L and SETD2 are two enzymes among others that catalyze H3K36 methylation. H3K4 methylation has also been reported for ASH1L. Radioactivity-based enzyme assays, Western and immunoblotting using specific antibodies and molecular modeling were used to characterize substrate specificity of ASH1L and SETD2. Here we report on the assay development and kinetic characterization of ASH1L and SETD2 and their substrate specificities in vitro. Both enzymes were active with recombinant nucleosome as substrate. However, SETD2 but not ASH1L methylated histone peptides as well indicating that the interaction of the basic post-SET domain region with substrate may not be critical for SETD2 activity. Both enzymes were not active with nucleosome containing a H3K36A mutation indicating their specificity for H3K36. Analyzing the methylation state of the products of ASH1L and SETD2 reactions also confirmed that both enzymes mono- and dimethylate H3K36 and are inactive with H3K4 as substrate, and that only SETD2 is able to trimethylate H3K36 in vitro. We determined the kinetic parameters for ASH1L and SETD2 activity enabling screening for inhibitors that can be used to further investigate the roles of these two proteins in health and disease. Both ASH1L and SETD2 are H3K36 specific methyltransferases but only SETD2 can trimethylate this mark. The basic post-SET domain region is critical for ASH1L but not SETD2 activity. We provide full kinetic characterization of ASH1L and SETD2 activity. Copyright © 2015. Published by Elsevier B.V.
    Biochimica et Biophysica Acta 05/2015; 1850(9). DOI:10.1016/j.bbagen.2015.05.013 · 4.66 Impact Factor
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