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|Title:||Identification of a cis-acting element in the rat α-fetoprotein gene and its specific binding proteins in F9 cells during retinoic acid-induced differentiation|
|Citation:||Chen, H.,Dong, J.-M.,Liu, Y.,Chiu, J.-F. (1999-01-01). Identification of a cis-acting element in the rat α-fetoprotein gene and its specific binding proteins in F9 cells during retinoic acid-induced differentiation. Journal of Cellular Biochemistry 72 (1) : 25-34. ScholarBank@NUS Repository. https://doi.org/10.1002/(SICI)1097-4644(19990101)72:13.0.CO;2-W|
|Abstract:||Mouse F9 embryonic teratocarcinoma stem cells can be induced to differentiate into visceral endoderm. Following retinoic acid (RA) treatment, alpha-fetoprotein (AFP), a differentiation marker, is expressed and secreted. The mechanism by which RA regulates AFP expression during differentiation is not clear. The relatively late induction of AFP indicates that the AFP gene may not be a primary target of RA activity during F9 cell differentiation. In this study, a CAT reporter plasmid containing the rat AFP 5'-regulatory region (-7040 to +7) adjacent to the CAT gene (pAFPCAT) was stably transfected into F9 cells and used to delineate a cis-acting element which associates with AFP gene activation. Similar spatial and temporal expression patterns between the transcriptional activity of the recombinant AFP gene and the endogenous AFP gene demonstrate that this stably transfected F9 system can be used to dissect both cis-elements and trans-acting factors responsible for RA-induced AFP expression. Using a series of deletion mutants of the pAFPCAT, the region between -2611 to -1855 was found to be important in AFP- induction. Subsequent analysis identified a functional sequence (-1905 to - 1891, 5'-ACTAAAATGGAGACT-3') that differentially binds nuclear proteins from undifferentiated and differentiated F9 cells. This sequence, designed as differentiation-associated sequence (DAS) for its unique binding of a nuclear protein (DAP-II) that appears during RA-induced F9 differentiation, acts as a regulatory protein factor in AFP gene activation.|
|Source Title:||Journal of Cellular Biochemistry|
|Appears in Collections:||Staff Publications|
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