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Hematopoietic stem cell differentiation involves the silencing of self-renewal genes and induction of a specific transcriptional program. Identification of multiple covalent cytosine modifications raises the question of how these derivatized bases influence stem cell commitment. Using a replicative primary human hematopoietic stem/progenitor cell differentiation system, we demonstrate dynamic changes of 5-hydroxymethylcytosine (5-hmC) during stem cell commitment and differentiation to the erythroid lineage. Genomic loci that maintain or gain 5-hmC density throughout erythroid differentiation contain binding sites for erythroid transcription factors and several factors not previously recognized as erythroid-specific factors. The functional importance of 5-hmC was demonstrated by impaired erythroid differentiation, with augmentation of myeloid potential, and disrupted 5-hmC patterning in leukemia patient-derived CD34+ stem/early progenitor cells with TET methylcytosine dioxygenase 2 (TET2) mutations. Thus, chemical conjugation and affinity purification of 5-hmC-enriched sequences followed by sequencing serve as resources for deciphering functional implications for gene expression during stem cell commitment and differentiation along a particular lineage.

Original publication

DOI

10.1016/j.celrep.2013.11.044

Type

Journal article

Journal

Cell Rep

Publication Date

16/01/2014

Volume

6

Pages

231 - 244

Keywords

5-Methylcytosine, Antigens, CD34, Cells, Cultured, Cytosine, DNA Methylation, Dioxygenases, Erythroid Cells, Erythropoiesis, Histones, Humans, Mutation, Regulatory Sequences, Nucleic Acid, Transcription Factors