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Identification and characterization of promoter and regulatory regions for mouse Adam2 gene expression

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Abstract

ADAM2, a member of the ‘a disintegrin and metalloprotease’ (ADAM) family, is a key protein in mammalian fertilization that is specifically expressed in testicular germ cells. Here, we investigated the transcriptional regulation of the mouse Adam2 gene. An in silico analysis identified two conserved non-coding sequences located upstream of the mouse and human ADAM2 genes. The upstream region of the mouse Adam2 gene was found to lack typical TATA and CAAT boxes, and to have a high GC content. Our in vitro transient transfection-reporter analysis identified a promoter in this region of the mouse Adam2 gene, along with regulatory regions that inhibit the activity of this promoter in somatic cells. Site-directed mutagenesis revealed that the caudal-type homeobox 1 and CCTC-binding factor motifs are responsible for the inhibitory activities of the repressor regions. Finally, electrophoretic mobility shift assays showed putative transcription factor–promoter DNA complexes, and DNA-affinity chromatography and proteomic analyses identified myelin gene regulatory factor as a binding partner of the Adam2 promoter. This provides the first identification and characterization of promoter and repressor regions that regulate the transcription of the mouse Adam2 gene, and offers insights into the regulation of this germ-cell-specific gene.

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Acknowledgments

This work was supported by the Korea Science and Engineering Foundation (Grant 2010-0028776), the Korea Research Foundation (Grant KRF-2008-313-C00736), and a Gwangju Institute of Science and Technology Systems Biology Infrastructure Establishment Grant.

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Authors and Affiliations

  1. School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, 500-712, Korea

    Heejin Choi, Boyeon Lee, Sora Jin, Jun Tae Kwon, Jihye Kim, Juri Jeong, Seungmin Oh, Zee Yong Park & Chunghee Cho

  2. Department of Life Science, The Catholic University of Korea, Bucheon, 420-743, Korea

    Byung-Nam Cho

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  1. Heejin Choi
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Correspondence to Chunghee Cho.

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Choi, H., Lee, B., Jin, S. et al. Identification and characterization of promoter and regulatory regions for mouse Adam2 gene expression. Mol Biol Rep 40, 787–796 (2013). https://doi.org/10.1007/s11033-012-2116-8

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  • DOI: https://doi.org/10.1007/s11033-012-2116-8

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