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J. Biol. Chem., Vol. 277, Issue 2, 1619-1627, January 11, 2002
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From the Department of Psychiatry and Behavioral Science, State
University of New York, Stony Brook, New York 11794-8101
Transcription from the amyloid precursor protein
(APP) promoter is largely dependent on a nuclear factor binding site
designated as APB
. The protein that binds to this site is the
multifunctional transcription factor CTCF, which consists of 727 amino
acids and contains a domain of 11 zinc finger motifs that is flanked by 267 amino acids on the N-terminal side and 150 amino acids on the
C-terminal side. Depleting HeLa cell nuclear extract of endogenous CTCF
specifically reduced transcriptional activity from the APP promoter. However, transcriptional activity was restored by
replenishing the depleted extract with recombinant CTCF. Deleting 201 amino acids from the C-terminal end of CTCF had no detrimental effect on transcriptional activation, whereas deleting either 248 or 284 amino
acids from the N-terminal end abolished transcriptional activation.
Competing endogenous CTCF in vivo was accomplished by
cotransfecting COS-1 cells with a plasmid overexpressing CTCF constructs and a reporter plasmid containing the APP
promoter. Under these conditions, an N-terminal deletion of CTCF
reduced expression from the APP promoter, whereas the
C-terminal deletion had no effect. These results demonstrate that CTCF
activates transcription from the APP promoter and that the
activation domain is located on the N-terminal side of the zinc finger domain.
To whom correspondence should be addressed. Tel.: 631-444-8025;
Fax: 631-444-7534; E-mail:
wquitschke@mail.psychiatry.sunysb.edu.
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