cAMP motifs regulating transcription in the aquaporin 2 gene

被引:138
作者
Hozawa, S
Holtzman, EJ
Ausiello, DA
机构
[1] MASSACHUSETTS GEN HOSP, RENAL UNIT, BOSTON, MA 02114 USA
[2] MASSACHUSETTS GEN HOSP, DEPT MED, BOSTON, MA 02114 USA
[3] HARVARD UNIV, SCH MED, BOSTON, MA 02114 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 1996年 / 270卷 / 06期
关键词
aquaporin 2 gene promoter; adenosine; 3'; 5'-cyclic monophosphate; gene transcription;
D O I
10.1152/ajpcell.1996.270.6.C1695
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Genomic clones including the 5' flanking regions of the AQP2 (aquaporin 2) gene were isolated, and the promoter region was examined by transiently transfecting a promoter-luciferase reporter fusion gene into renal cultured epithelial cells. An orientation specific promoter for the AQP2 gene was found within the proximal 3 kb of 5'-flanking region. Minimal basal promoter activity of the AQP2 gene was found within 198 bp upstream from the transcription start site by deletion analysis. Sequencing the transcriptionally active region revealed a typical TATA box, adenosine 3',5'-cyclic monophosphate (cAMP) responsive element (CRE) and three putative CCAAT boxes in the proximal 1.2-kb region. Significantly, a GATA motif, AP1, AP2, and SP1 transcriptional factor consensus sites were also found in this region. Exposure to cAMP-enhancing agents (1 nM vasopressin or 20 mM forskolin and 250 mM 3-isobutyl-1-methylxanthine) showed that these agents increased luciferase activity in a parallel fashion, suggesting that vasopressin-induced AQP2 gene transcription is mediated through increases in intracellular cAMP in at least one renal cell type, the LLC-PK1 cells. The mechanism of cAMP responsiveness of AQP2 gene transcription was further studied using a series of deletion mutants in renal epithelial cells and other cell types. The cAMP regulatory motifs were shown to exist in a 50-bp sequence between -340 and -290 (containing CRE) and a 65-bp sequence (containing an AP2 site) between -150 and the ATG start site in LLC-PK1 cells. In rat inner medullary collecting duct (IMCD) cells, the cAMP regulatory motifs also exist in a 50-bp sequence between -340 and -290 (containing CRE) and in a 10-bp sequence between -160 and -150 (containing an SP1 site). These separate regions may cooperate to confer full cAMP inducibility to the AQP2 gene in a cell-specific manner.
引用
收藏
页码:C1695 / C1702
页数:8
相关论文
共 29 条
[1]   AMPLIFICATION OF THE TRANSCRIPTIONAL SIGNAL MEDIATED BY THE TANDEM CAMP RESPONSE ELEMENTS OF THE GLYCOPROTEIN HORMONE ALPHA-SUBUNIT GENE OCCURS THROUGH SEVERAL DISTINCT MECHANISMS [J].
ANDERSEN, B ;
KENNEDY, GC ;
HAMERNIK, DL ;
BOKAR, JA ;
BOHINSKI, R ;
NILSON, JH .
MOLECULAR ENDOCRINOLOGY, 1990, 4 (04) :573-582
[2]  
AUSUBEL F. M., 1992, CURRENT PROTOCOL MOL, P4757
[3]   CHARACTERIZATION OF THE HUMAN TRYPTOPHAN-HYDROXYLASE GENE PROMOTER - TRANSCRIPTIONAL REGULATION BY CAMP REQUIRES A NEW MOTIF DISTINCT FROM THE CAMP-RESPONSIVE ELEMENT [J].
BOULARAND, S ;
DARMON, MC ;
RAVASSARD, P ;
MALLET, J .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (08) :3757-3764
[4]  
BRASIER AR, 1989, BIOTECHNIQUES, V7, P1116
[5]   MEMBRANE RECYCLING AND EPITHELIAL-CELL FUNCTION [J].
BROWN, D .
AMERICAN JOURNAL OF PHYSIOLOGY, 1989, 256 (01) :F1-F12
[6]  
BUSTROS A, 1992, BIOCHEM BIOPH RES CO, V189, P1157
[7]   A CYCLIC AMP- AND PHORBOL ESTER-INDUCIBLE DNA ELEMENT [J].
COMB, M ;
BIRNBERG, NC ;
SEASHOLTZ, A ;
HERBERT, E ;
GOODMAN, HM .
NATURE, 1986, 323 (6086) :353-356
[8]   REQUIREMENT OF HUMAN RENAL WATER CHANNEL AQUAPORIN-2 FOR VASOPRESSIN-DEPENDENT CONCENTRATION OF URINE [J].
DEEN, PMT ;
VERDIJK, MAJ ;
KNOERS, NVAM ;
WIERINGA, B ;
MONNENS, LAH ;
VANOS, CH ;
VANOOST, BA .
SCIENCE, 1994, 264 (5155) :92-95
[9]   MEMBRANE LOCALIZATION OF THE PERTUSSIS TOXIN-SENSITIVE G-PROTEIN SUBUNIT-ALPHA-I-2 AND SUBUNIT-ALPHA-I-3 AND EXPRESSION OF A METALLOTHIONEIN-ALPHA-I-2 FUSION GENE IN LLC-PK1 CELLS [J].
ERCOLANI, L ;
STOW, JL ;
BOYLE, JF ;
HOLTZMAN, EJ ;
LIN, H ;
GROVE, JR ;
AUSIELLO, DA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (12) :4635-4639
[10]   CLONING AND EXPRESSION OF APICAL MEMBRANE WATER CHANNEL OF RAT-KIDNEY COLLECTING TUBULE [J].
FUSHIMI, K ;
UCHIDA, S ;
HARA, Y ;
HIRATA, Y ;
MARUMO, F ;
SASAKI, S .
NATURE, 1993, 361 (6412) :549-552