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Influence of Repressive Histone and DNA Methylation upon D4Z4 Transcription in Non-Myogenic Cells.

Title: Influence of Repressive Histone and DNA Methylation upon D4Z4 Transcription in Non-Myogenic Cells.
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Name(s): Das, Sunny, author
Chadwick, Brian P, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-07-28
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: We looked at a disease-associated macrosatellite array D4Z4 and focused on epigenetic factors influencing its chromatin state outside of the disease-context. We used the HCT116 cell line that contains the non-canonical polyadenylation (poly-A) signal required to stabilize somatic transcripts of the human double homeobox gene DUX4, encoded from D4Z4. In HCT116, D4Z4 is packaged into constitutive heterochromatin, characterized by DNA methylation and histone H3 tri-methylation at lysine 9 (H3K9me3), resulting in low basal levels of D4Z4-derived transcripts. However, a double knockout (DKO) of DNA methyltransferase genes, DNMT1 and DNMT3B, but not either alone, results in significant loss of DNA and H3K9 methylation. This is coupled with upregulation of transcript levels from the array, including DUX4 isoforms (DUX4-fl) that are abnormally expressed in somatic muscle in the disease Facioscapulohumeral muscular dystrophy (FSHD) along with DUX4 protein, as indicated indirectly by upregulation of bondafide targets of DUX4 in DKO but not HCT116 cells. Results from treatment with a chemical inhibitor of histone methylation in HCT116 suggest that in the absence of DNA hypomethylation, H3K9me3 loss alone is sufficient to facilitate DUX4-fl transcription. Additionally, characterization of a cell line from a patient with Immunodeficiency, Centromeric instability and Facial anomalies syndrome 1 (ICF1) possessing a non-canonical poly-A signal and DNA hypomethylation at D4Z4 showed DUX4 target gene upregulation in the patient when compared to controls in spite of retention of H3K9me3. Taken together, these data suggest that both DNA methylation and H3K9me3 are determinants of D4Z4 silencing. Moreover, we show that in addition to testis, there is appreciable expression of spliced and polyadenylated D4Z4 derived transcripts that contain the complete DUX4 open reading frame (ORF) along with DUX4 target gene expression in the thymus, suggesting that DUX4 may provide normal function in this somatic tissue.
Identifier: FSU_pmch_27467759 (IID), 10.1371/journal.pone.0160022 (DOI), PMC4965136 (PMCID), 27467759 (RID), 27467759 (EID), PONE-D-15-54558 (PII)
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4965136.
Subject(s): Alleles
DNA Methylation
DNA Modification Methylases/genetics
HCT116 Cells
HEK293 Cells
Heterochromatin/metabolism
Histones/metabolism
Humans
Transcription, Genetic
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_27467759
Host Institution: FSU
Is Part Of: PloS one.
1932-6203
Issue: iss. 7, vol. 11

Choose the citation style.
Das, S., & Chadwick, B. P. (2016). Influence of Repressive Histone and DNA Methylation upon D4Z4 Transcription in Non-Myogenic Cells. Plos One. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_27467759