Transcriptional activation of the murine CTP:phosphocholine cytidylyltransferase gene (Ctpct):: combined action of upstream stimulatory and inhibitory cis-acting elements

被引:37
作者
Bakovic, M
Waite, K
Tang, W
Tabas, I
Vance, DE [1 ]
机构
[1] Univ Alberta, Dept Biochem, Edmonton, AB T6G 2S2, Canada
[2] Columbia Univ Coll Phys & Surg, Dept Med, New York, NY 10032 USA
[3] Columbia Univ Coll Phys & Surg, Dept Anat & Cell Biol, New York, NY 10032 USA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS | 1999年 / 1438卷 / 01期
基金
英国医学研究理事会;
关键词
phosphatidylcholine; CTP : phosphocholine cytidylyltransferase; ctpct promoter; gene regulation; nuclear factor; regulatory element;
D O I
10.1016/S1388-1981(99)00042-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
CTP:phosphocholine cytidylyltransferase plays a key role in regulating the rate of phosphatidylcholine biosynthesis. However, the proximal regulatory elements for the gene (Ctpct) that encode this enzyme and the cognate transcription factors involved have not been characterized. Ctpct promoter activities were deduced from promoter deletion constructs linked to a luciferase reporter and transiently transfected into C3H10T1/2 and McArdle RH7777 cells. Positive regulatory elements were located between -130 and -52 bp from the transcription start site. Basal expression resided downstream between -52 and +38 bp. DNase I protection and electromobility-shift assays indicated that Sp1-related nuclear factors bind to a stimulatory, a possible inhibitory and minimal promoter element. Gel-shift assays confirmed that all three regulatory regions bound Sp1. Sp1 was further implicated when Sp1-deficient Drosophila cells were co-transfected with promoter-reporter constructs and an Sp1 construct. DNase I assays also indicated that the Ap1 binding elements could be occupied in the proximal activator and minimal promoter regions. Gel-shift assays demonstrated that the distal activator region could bind Ap1 and an unknown transcription factor. We conclude that Sp1, Ap1 and an unknown transcription factor have important roles in regulating expression of the Ctpct gene. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:147 / 165
页数:19
相关论文
共 74 条
[1]   A RAPID MICROPREPARATION TECHNIQUE FOR EXTRACTION OF DNA-BINDING PROTEINS FROM LIMITING NUMBERS OF MAMMALIAN-CELLS [J].
ANDREWS, NC ;
FALLER, DV .
NUCLEIC ACIDS RESEARCH, 1991, 19 (09) :2499-2499
[2]   STEROL REGULATION OF FATTY-ACID SYNTHASE PROMOTER - COORDINATE FEEDBACK-REGULATION OF 2 MAJOR LIPID PATHWAYS [J].
BENNETT, MK ;
LOPEZ, JM ;
SANCHEZ, HB ;
OSBORNE, TF .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (43) :25578-25583
[3]   Transcriptional regulation of neuronal nicotinic acetylcholine receptor genes - Functional interactions between Sp1 and the rat beta 4 subunit gene promoter [J].
Bigger, CB ;
Casanova, EA ;
Gardner, PD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (51) :32842-32848
[4]  
Choi SB, 1997, MOL CELLS, V7, P58
[5]   TRANSLOCATION OF CTP-PHOSPHOCHOLINE CYTIDYLTRANSFERASE FROM CYTOSOL TO MEMBRANES IN HELA CELLS-STIMULATION BY FATTY-ACID, FATTY ALCOHOL, MONOACYGLYCEROL AND DIACYLGLYCEROL [J].
CORNELL, R ;
VANCE, DE .
BIOCHIMICA ET BIOPHYSICA ACTA, 1987, 919 (01) :26-36
[6]   ANALYSIS OF SP1 INVIVO REVEALS MULTIPLE TRANSCRIPTIONAL DOMAINS, INCLUDING A NOVEL GLUTAMINE-RICH ACTIVATION MOTIF [J].
COUREY, AJ ;
TJIAN, R .
CELL, 1988, 55 (05) :887-898
[7]   Inverse correlation between expression of phosphatidylethanolamine N-methyltransferase-2 and growth rate of perinatal rat livers [J].
Cui, Z ;
Shen, YJ ;
Vance, DE .
BIOCHIMICA ET BIOPHYSICA ACTA-LIPIDS AND LIPID METABOLISM, 1997, 1346 (01) :10-16
[8]   Expression of phosphatidylethanolamine N-methyltransferase-2 in McArdle-RH7777 hepatoma cells inhibits the CDP-choline pathway for phosphatidylcholine biosynthesis via decreased gene expression of CTP:phosphocholine cytidylyltransferase [J].
Cui, Z ;
Houweling, M ;
Vance, DE .
BIOCHEMICAL JOURNAL, 1995, 312 :939-945
[9]   SIMIAN VIRUS-40 EARLY PROMOTER MUTATIONS THAT AFFECT PROMOTER FUNCTION AND AUTO-REGULATION BY LARGE T-ANTIGEN [J].
DAS, GC ;
SALZMAN, NP .
JOURNAL OF MOLECULAR BIOLOGY, 1985, 182 (02) :229-239
[10]  
DAWSON PA, 1988, J BIOL CHEM, V263, P3372