One or more labile proteins regulate the stability of chimeric mRNA containing the 3′-untranslated region of cholesterol-7α-hydroxylase mRNA

被引:32
作者
Baker, DM [1 ]
Wang, SL [1 ]
Bell, DJ [1 ]
Drevon, CA [1 ]
Davis, RA [1 ]
机构
[1] San Diego State Univ, Dept Biol, Mammalian Cell & Mol Biol Lab, San Diego, CA 92182 USA
关键词
D O I
10.1074/jbc.M002351200
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
Multiple AUUUA elements similar to those that regulate the degradation of several different mRNAs are conserved in the 3'-untranslated region (3'-UTR) of cholesterol-7 alpha-hydroxylase (CYP7A1) mRNAs from several species. We examined if stabilization of mRNA decay could account for the >20-fold increase in the expression of CYP7A1 mRNA without a detectable change in transcription following dexamethasone treatment of rat hepatoma cells (L35 cells). Following RNA polymerase II-dependent transcription block or protein synthesis block, the decay of CYP7A1 mRNA displayed a short half-life (similar to 30 min). Control experiments showed that in cells pre-treated with a RNA polymerase II inhibitor, dexamethasone had no detectable effect on CYP7A1 mRNA decay, Stable expression of luciferase reporter mRNAs in L35 cells showed that the CYP7A1 3'-UTR was required to observe a dexamethasone induction. To examine the hypothesis that a labile protein is required for dexamethasone-induced mRNA stabilization, cells were stably transfected with a tetracycline-repressible promoter that drives the expression of a green fluorescent protein analogue (ECFP) with or without the 3'-UTR of CYP7A1, Cells expressing ECFP with the 5'-UTR of CYP7A1 displayed a 3-fold dexamethasone induction of ECFP mRNA, whereas cells expressing ECFP without the 3'-UTR did not. Moreover, specific block of the transcription of ECFP containing the 3'-UTR by adding the tetracycline analogue doxycycline clearly displayed dexamethasone-induced stabilization of mRNA decay, These data provide compelling evidence that a putative labile protein and the 3'-UTR of CYP7A1 act together to decrease the rate of CYP7A1 mRNA degradation.
引用
收藏
页码:19985 / 19991
页数:7
相关论文
共 49 条
[1]  
Agellon LB, 1997, BIOCHEM J, V328, P393
[2]   Analysis of human CYP7A1 mRNA decay in HepG2 cells by reverse transcription-polymerase chain reaction [J].
Andreou, ER ;
Prokipcak, RD .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1998, 357 (01) :137-146
[3]  
CHEN CYA, 1995, MOL CELL BIOL, V15, P5777
[4]   SELECTIVE DEGRADATION OF EARLY-RESPONSE-GENE MESSENGER-RNAS - FUNCTIONAL ANALYSES OF SEQUENCE FEATURES OF THE AU-RICH ELEMENTS [J].
CHEN, CYA ;
SHYU, AB .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (12) :8471-8482
[5]   2 CELLULAR PROTEINS BIND SPECIFICALLY TO A PURINE-RICH SEQUENCE NECESSARY FOR THE DESTABILIZATION FUNCTION OF A C-FOS PROTEIN-CODING REGION DETERMINANT OF MESSENGER-RNA INSTABILITY [J].
CHEN, CYA ;
YOU, Y ;
SHYU, AB .
MOLECULAR AND CELLULAR BIOLOGY, 1992, 12 (12) :5748-5757
[6]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[7]   GENOMIC CLONING, SEQUENCING, AND ANALYSIS OF THE HAMSTER CHOLESTEROL 7-ALPHA-HYDROXYLASE GENE (CYP7) [J].
CRESTANI, M ;
GALLI, G ;
CHIANG, JYL .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1993, 306 (02) :451-460
[8]  
EDWARDS PA, 1996, N COMP BIOC, V31, P341
[9]  
Feingold KR, 1996, J LIPID RES, V37, P223
[10]   Basic transcription element binding protein (BTEB) transactivates the cholesterol 7α-hydroxylase gene (CYP7A) [J].
Foti, D ;
Stroup, D ;
Chiang, JYL .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 253 (01) :109-113