Nucleic Acids Research, Vol 26, Issue 7 1801-1806, Copyright © 1998 by Oxford University Press
MP Clark, CW Chow, JE Rinaldo and R Chalkley
In the present study, we have shown that a downstream element located in
the coding region of the TATA-less rat xanthine dehydrogenase/oxidase
(XDH/XO) gene (-7 to +42) plays an important role in transcription
initiation and C/EBP transcriptional activation. Previous work from our
laboratory has shown that the promoter is organized with multiple initiator
elements (Inr 1, 2, 3 and 4) which are important for transcription
initiation. Additionally, we had identified two C/EBP binding sites
upstream of this promoter. Deletional and mutational studies revealed that
C/EBP binding was not essential for the basal level of transcriptional
initation. However when XO-luciferase constructs include downstream
sequence extending to +42 there is development of C/EBP sensitivity as well
as a shift in the initiator usage. In the absence of the downstream
element, primer extension analyses reveals Inr 3 and 4 to be the major
start sites but in the presence of this additional sequence the usage is
shifted to Inr 1 and 2. This shift in Inr usage more closely resembles that
seen in intact macrophages or liver cells. Gel mobility shift assays
indicate the presence of several binding factors located in this downstream
region, one of which has been identified as YY-1. We postulate that YY- 1
allows DNA bending which permits the upstream C/EBP elements to exhibit a
transcriptional activation which is not seen when the downstream element is
absent. This study presents a potential model for regulation of the XDH/XO
promoter.
ARTICLES
Correct usage of multiple transcription initiation sites and C/EBP- dependent transcription activation of the rat XDH/XO TATA-less promoter requires downstream elements located in the coding region of the gene
Department of Medicine, School of Medicine, Vanderbilt University, Nashville, TN 37232, USA.
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