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https://hdl.handle.net/2440/27599
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Type: | Journal article |
Title: | Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in β-cells |
Other Titles: | Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in beta-cells |
Author: | Sunyakumthorn, P. Boonsaen, T. Boonsaeng, V. Wallace, J. Jitrapakdee, S. |
Citation: | Biochemical and Biophysical Research Communications, 2005; 329(1):188-196 |
Publisher: | Academic Press Inc |
Issue Date: | 2005 |
ISSN: | 0006-291X |
Statement of Responsibility: | Piyanate Sunyakumthorn, Thirajit Boonsaen, Vichai Boonsaeng, John C. Wallace and Sarawut Jitrapakdee |
Abstract: | Pyruvate carboxylase plays diverse roles in different biosynthetic pathways, including glucose-induced insulin secretion in pancreatic β-cells. We have localized the control region of the P2 promoter by generating a series of 5′-nested deletion constructs, and both 25- and 9-bp internal deletion constructs, as well as by performing site-directed mutagenesis. Transient transfections of these constructs into INS-1 cells identified a CCAAT box and a GC box that are located at −65/−61 and −48/−41, respectively, as the important determinants. Disruption of the GC box resulted in a 4-fold reduction of the reporter activity, while disruption of the proximal CCAAT box (−65/−61) but not the distal CCAAT box (−95/−91) increased the reporter activity by 3-fold. Simultaneous disruptions of both the GC box and the CCAAT box reduced the reporter activity to a level that was close to that of the single GC box mutation. Electrophoretic mobility shift assays (EMSAs) and supershift EMSAs using nuclear extract from INS-1 cells demonstrated that Sp1 and Sp3 bind a GC box while the nuclear factor Y was shown to bind the proximal but not the distal CCAAT box. |
Keywords: | Pyruvate carboxylase Biotin enzyme Glucose-induced insulin release Gene transcription Sp1 Sp3 NF-Y |
Rights: | © 2005 Elsevier Inc. All rights reserved. |
DOI: | 10.1016/j.bbrc.2005.01.108 |
Published version: | http://dx.doi.org/10.1016/j.bbrc.2005.01.108 |
Appears in Collections: | Aurora harvest 2 Molecular and Biomedical Science publications |
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