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The CCAAT element in the CIWI promoter regulates transcriptional initiation in chicken primordial germ cells

Cited 4 time in Web of Science Cited 4 time in Scopus
Authors
Sohn, Yoon Ah; Lee, Sang In; Choi, Hee Jung; Kim, Hyun Jeong; Kim, Ki Hyun; Park, Tae Sub; Han, Jae Yong
Issue Date
2014
Publisher
Wiley
Citation
Molecular Reproduction and Development, vol.81 no.9, pp. 871-882
Abstract
The P-element-induced wimpy testis (PIWI) protein, which associates with small non-coding RNAs, is responsible for maintaining the integrity of germ cells and stem cells. Thus, transcriptional regulation of PIWI is critical for its effective functional modulation. In this study, we identified the promoter region of the PIWI homolog in chicken (CIWI), and investigated the transcriptional regulatory elements that control expression of CIWI in chicken primordial germ cells (PGCs). We constructed a vector that included the enhanced green fluorescent protein (eGFP) gene controlled by the 4-kb CIWI promoter. The vector was expressed in chicken PGCs, but not in chicken embryonic fibroblasts. Based on promoter deletion and fragmentation assays, we found that a 252-bp fragment of the CIWI promoter (−577 to −326 bp) was crucial for CIWI expression in PGCs. A CCAAT transcriptional regulatory element (−498 to −494 bp) was detected in the proximal region from the transcription initiation site of CIWI, and mutational analysis confirmed that this element regulates transcriptional initiation in chicken PGCs. Interestingly, the regions flanking the CCAAT element, which are positioned differently in HIWI (human), Miwi (mouse), and CIWI orthologs, were highly conserved. In addition, we predicted that specificity protein 1 (SP1) motifs modulate the transcriptional initiation of CIWI by binding to the 5'-flanking regions of the CCAAT box. Overall, 252 bp of the CIWI promoter possessing the transcriptional regulatory element CCAAT is crucial for regulating CIWI gene expression in chicken PGCs. This promoter may be applicable for the regulation of CIWI expression during germ-cell development.
ISSN
1040-452X
Language
English
URI
https://hdl.handle.net/10371/100379
DOI
https://doi.org/10.1002/mrd.22356
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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