Exp Mol Med.
2005 Dec;37(6):601-607.
Identification and functional characterization of an alternative splice variant within the fourth exon of human nanog
- Affiliations
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- 1Division of Molecular and Life Sciences, College of Science and Technology, Hanyang University, Seoul 133-791, Korea. iychu@hanyang.ac.kr
- 2Department of Microbiology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
- 3Department of Internal Medicine, Hanyang University Hospital, Seoul 133-792, Korea.
- 4Genome Research Center for Asthma and Allergic Diseases, Division of Allergy and Respiratory Medicine, Soonchunhyang University Hospital, Seoul and Bucheon, Korea.
Abstract
- Nanog, a homeodomain (HD) transcription factor, plays a critical role in the maintenance of embryonic stem (ES) cell self-renewal. Here, we report the identification of an alternatively-spliced variant of nanog. This variant lacked a stretch of amino acids (residues 168-183) located between the HD and tryptophan-repeat (WR) of the previously-reported full length sequence, suggesting that the deleted sequence functions as a linker and possibly affects the flexibility of the C-terminal transactivation domain relative to the DNA binding domain. Expression of mRNA encoding the splice variant, designated as nanog-delta 48, was much lower than that of the full length version in human ES cells. The ratio of nanog-delta 48 transcript to full length transcript increased, however, in multipotent adult progenitor cells. EMSA analysis revealed that both forms of Nanog were able to bind a nanog binding sequence with roughly the same affinity. A reporter plasmid assay also showed that both variants of nanog modestly repressed transactivation of gata-4, whose expression is proposed to be inhibited by nanog, with comparable potency. We conclude that, despite the difference in primary structure and expression pattern in various stem cells, the alternatively-spliced variant of Nanog has similar activity to that of the full length version.
Keyword
MeSH Terms
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Alternative Splicing/*genetics
Amino Acid Sequence
Base Sequence
Cell Nucleus
Cells, Cultured
DNA-Binding Proteins/chemistry/*genetics/metabolism
Exons/*genetics
GATA4 Transcription Factor/metabolism
Gene Expression Profiling
Genes, Reporter
Homeodomain Proteins/chemistry/*genetics/metabolism
Humans
Introns/genetics
Molecular Sequence Data
Promoter Regions (Genetics)
RNA, Messenger/genetics/metabolism
Research Support, Non-U.S. Gov't
Trans-Activation (Genetics)
Transfection
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