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, 8 (12), 5179-87

Interaction of a Hepatoma-Specific Nuclear Factor With Transcription-Regulatory Sequences of the Human Alpha-Fetoprotein and Albumin Genes

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Interaction of a Hepatoma-Specific Nuclear Factor With Transcription-Regulatory Sequences of the Human Alpha-Fetoprotein and Albumin Genes

K Sawadaishi et al. Mol Cell Biol.

Abstract

DNase I footprinting and gel mobility shift analysis showed that an HuH-7 hepatoma nuclear protein, termed AFP1, binds specifically to an AT-rich sequence, TGATTAATAATTACA, in domain B of the human alpha-fetoprotein enhancer. No such binding activity was found in HeLa cell nuclei. Transient transfection studies showed that a 54-base-pair region corresponding to the AFP1-binding site could stimulate the simian virus 40 early promoter to express a linked chloramphenicol acetyltransferase gene in an orientation-independent and cell-specific manner. The correlation between the binding of AFP1 and the stimulation of chloramphenicol acetyltransferase gene expression strongly suggests that specific interaction of AFP1 with the AT motif is important for cell-specific transcriptional enhancement. Competition gel mobility shift analysis revealed that similar AT-rich sequences with high affinities to AFP1 were also present in the promoters of the alpha-fetoprotein and albumin genes. These results suggest that AFP1 may function as a common regulatory factor in the transcription of the alpha-fetoprotein and albumin genes.

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