TY - JOUR
T1 - The human GATA1 gene retains a 5= insulator that maintains chromosomal architecture and GATA1 expression levels in splenic erythroblasts
AU - Moriguchi, Takashi
AU - Yu, Lei
AU - Takai, Jun
AU - Hayashi, Makiko
AU - Satoh, Hironori
AU - Suzuki, Mikiko
AU - Ohneda, Kinuko
AU - Yamamoto, Masayuki
N1 - Publisher Copyright:
© 2015, American Society for Microbiology.
PY - 2015
Y1 - 2015
N2 - GATA1 is a key transcription factor for erythropoiesis. GATA1 gene expression is strictly regulated at the transcriptional level. While the regulatory mechanisms governing mouse Gata1 (mGata1) gene expression have been studied extensively, how expression of the human GATA1 (hGATA1) gene is regulated remains to be elucidated. To address this issue, we generated hGATA1 bacterial artificial chromosome (BAC) transgenic mouse lines harboring a 183-kb hGATA1 locus covering the hGATA1 exons and distal flanking sequences. Transgenic hGATA1 expression coincides with endogenous mGata1 expression and fully rescues hematopoietic deficiency in mGata1 knockdown mice. The transgene exhibited copy number-dependent and integration position- independent expression of hGATA1, indicating the presence of chromatin insulator activity within the transgene. We found a novel insulator element at 29 kb 5= to the hGATA1 gene and refer to this element as the 5= CCCTC-binding factor (CTCF) site. Substitution mutation of the 5= CTCF site in the hGATA1 BAC disrupted the chromatin architecture and led to a reduction of hGATA1 expression in splenic erythroblasts under conditions of stress erythropoiesis. Our results demonstrate that expression of the hGATA1 gene is regulated through the chromatin architecture organized by 5= CTCF site-mediated intrachromosomal interactions in the hGATA1 locus.
AB - GATA1 is a key transcription factor for erythropoiesis. GATA1 gene expression is strictly regulated at the transcriptional level. While the regulatory mechanisms governing mouse Gata1 (mGata1) gene expression have been studied extensively, how expression of the human GATA1 (hGATA1) gene is regulated remains to be elucidated. To address this issue, we generated hGATA1 bacterial artificial chromosome (BAC) transgenic mouse lines harboring a 183-kb hGATA1 locus covering the hGATA1 exons and distal flanking sequences. Transgenic hGATA1 expression coincides with endogenous mGata1 expression and fully rescues hematopoietic deficiency in mGata1 knockdown mice. The transgene exhibited copy number-dependent and integration position- independent expression of hGATA1, indicating the presence of chromatin insulator activity within the transgene. We found a novel insulator element at 29 kb 5= to the hGATA1 gene and refer to this element as the 5= CCCTC-binding factor (CTCF) site. Substitution mutation of the 5= CTCF site in the hGATA1 BAC disrupted the chromatin architecture and led to a reduction of hGATA1 expression in splenic erythroblasts under conditions of stress erythropoiesis. Our results demonstrate that expression of the hGATA1 gene is regulated through the chromatin architecture organized by 5= CTCF site-mediated intrachromosomal interactions in the hGATA1 locus.
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U2 - 10.1128/MCB.00011-15
DO - 10.1128/MCB.00011-15
M3 - Article
C2 - 25755285
AN - SCOPUS:84929207965
VL - 35
SP - 1825
EP - 1837
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
SN - 0270-7306
IS - 10
ER -