IDENTIFICATION OF A CIS-ACTING ELEMENT IN THE RAT ALPHA-FETOPROTEIN GENE AND ITS SPECIFIC BINDING-PROTEINS IN F9 CELLS DURING RETINOIC ACID-INDUCED DIFFERENTIATION

Citation
Hm. Chen et al., IDENTIFICATION OF A CIS-ACTING ELEMENT IN THE RAT ALPHA-FETOPROTEIN GENE AND ITS SPECIFIC BINDING-PROTEINS IN F9 CELLS DURING RETINOIC ACID-INDUCED DIFFERENTIATION, Journal of cellular biochemistry, 72(1), 1999, pp. 25-34
Citations number
39
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,"Cell Biology
ISSN journal
0730-2312
Volume
72
Issue
1
Year of publication
1999
Pages
25 - 34
Database
ISI
SICI code
0730-2312(1999)72:1<25:IOACEI>2.0.ZU;2-9
Abstract
Mouse F9 embryonic teratocarcinoma stem cells can be induced to differ entiate into visceral endoderm. Following retinoic acid (RA) treatment , alpha-fetoprotein (AFP), a differentiation marker is expressed and s ecreted. The mechanism by which RA regulates AFP expression during dif ferentiation is not clear. The relatively late induction of AFP indica tes that the AFP gene may not be a primary target of RA activity durin g F9 cell differentiation. In this study, a CAT reporter plasmid conta ining the rat AFP 5'-regulatory region (- 7040 to + 7) adjacent to the CAT gene (pAFPCAT) was stably transfected into F9 cells and used to d elineate a cis-acting element which associates with AFP gene activatio n. Similar spatial and temporal expression patterns between the transc riptional 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 pAF PCAT, the region between -2611 to -1855 was found to be important in A FP-induction. Subsequent analysis identified a functional sequence (-1 905 to -1891, 5'-ACTAAAATGGAGACT-3') that differentially binds nuclear proteins from undifferentiated and differentiated F9 cells. This sequ ence, designed as differentiation-associated sequence (DAS) for its un ique binding of a nuclear protein (DAP-II) that appears during RA-indu ced F9 differentiation, acts as a regulatory protein factor in AFP gen e activation. J. Cell. Biochem. 72:25-34, 1999. (C) 1999 Wiley-Liss, I nc.