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Characterisation of histone variant distribution in human embryonic stem cells by transfection of in vitro transcribed mRNA
Author(s) -
Shaw Margaret L.,
Williams Evan J.,
Hawes Susan,
Saffery Richard
Publication year - 2009
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.21077
Subject(s) - biology , chromatin , embryonic stem cell , histone , microbiology and biotechnology , induced pluripotent stem cell , histone h3 , subcellular localization , histone h1 , transfection , stem cell , gene , genetics
Abstract Recent studies, primarily in mouse embryonic stem cells, have highlighted the unique chromatin state of pluripotent stem cells, including the incorporation of histone variants into specific genomic locations, and its role in facilitating faithful expression of genes during development. However, there is little information available on the expression and subcellular localisation of histone variants in human embryonic stem cells (hESCs). In this study, we confirmed the expression of a panel of histone variant genes in several hESC lines and demonstrated the utility of transfection of in vitro transcribed, epitope‐tagged mRNAs to characterise the subcellular localisation of these proteins. The subcellular localisations of variant histone H3 (CENP‐A, H3.3), H2A (MACROH2A, H2AX, H2AZ, H2ABBD) and H1 (H1A, HB, H1C, H1D) were examined, revealing distinct nuclear localisation profiles for each protein. These data highlight the differences between murine (m) ESCs and hESCs, including the presence of a MACROH2A‐enriched inactive X chromosome in undifferentiated XX hESC lines. We also provide the first evidence for MACROH2A accumulation on the Y‐chromosome in XY hESCs. Mol. Reprod. Dev. 76: 1128–1142, 2009. © 2009 Wiley‐Liss, Inc.