Creb and Sp/Kruppel response elements cooperate to control rat TRH gene transcription in response to cAMP

被引:11
作者
Cote-Velez, Antonieta [1 ]
Perez-Maldonado, Adrian [1 ]
Osuna, Joel [2 ]
Barrera, Benito [1 ]
Charli, Jean-Louis [1 ]
Joseph-Bravo, Patricia [1 ]
机构
[1] UNAM, Dept Genet Desarrollo & Fisiol Mol, Inst Biotecnol, Cuernavaca 62271, Morelos, Mexico
[2] UNAM, Dept Ingn Celular & Biocatalisis, Inst Biotecnol, Cuernavaca 62271, Morelos, Mexico
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS | 2011年 / 1809卷 / 03期
关键词
TRH; CRE; cAMP; Sp/KFL; Hypothalamus; Transcription; THYROTROPIN-RELEASING-HORMONE; RNA-POLYMERASE-II; THYROID-HORMONE; PARAVENTRICULAR NUCLEUS; BINDING PROTEIN; MESSENGER-RNA; COLD-EXPOSURE; HYPOTHALAMIC NEURONS; TRHERGIC NEURONS; PROMOTER;
D O I
10.1016/j.bbagrm.2011.01.005
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
Expression of hypophysiotropic TRH, that controls thyroid axis activity, is increased by cold exposure; this effect is mimicked in rat hypothalamic cells incubated with norepinephrine or cAMP analogs. TRH proximal promoter contains three putative CRE: Site-4 or CRE-1 that overlaps an element recognized by thyroid hormone receptors, CRE-2 with adjacent sequences GC box or CACCC recognized by Sp/Kruppel factors (extended CRE-2), and AP-1 sites flanking a GRE(1/2). To evaluate the role of each element in the cAMP response, these sites were mutated or deleted in rat TRH promoter linked to luciferase gene (TRH-luc) and co-transfected with beta-gal expression vector in various cell lines; C6 cells gave the highest response to forskolin. Basal activity was most affected by mutations or deletion of CRE-2 site, or CACCC (50-75% of wild type-WT). Forskolin-induced 3x stimulation in WT which decreased 25% with CRE-1 or AP-1 deletions, but 50% when CRE-2 or its 5' adjacent GC box was altered. SH-SY5Y cells co-transfected with CREB-expression vector increased dB-cAMP response in the wild type but not in the CRE-2 mutated plasmid; cotransfecting CREB-A (a dominant negative expression vector) strongly diminished basal or cAMP response. Primary cultures of hypothalamic cells transfected with plasmids containing deletions of CRE-1, CRE-2, or extended CRE-2 failed to respond to forskolin when CRE-2 was modified. These results corroborate the CRE-2 site as the main cAMP-response element of rat TRH promoter, not exclusive of transcription factors of hypothalamic cells, and stress the relevance of adjacent Sp-1 sites, important mediators of some metabolic hormones. Published by Elsevier B.V.
引用
收藏
页码:191 / 199
页数:9
相关论文
共 56 条
[1]   The expression of TRH, its receptors and degrading enzyme is differentially modulated in the rat limbic system during training in the Morris water maze [J].
Aguilar-Valles, Argel ;
Sanchez, Edith ;
de Gortari, Patricia ;
Garcia-Vazquez, Arlene I. ;
Ramirez-Amaya, Victor ;
Bermudez-Rattoni, Federico ;
Joseph-Bravo, Patricia .
NEUROCHEMISTRY INTERNATIONAL, 2007, 50 (02) :404-417
[2]   A dominant-negative inhibitor of CREB reveals that it is a general mediator of stimulus-dependent transcription of c-fos [J].
Ahn, S ;
Olive, M ;
Aggarwal, S ;
Krylov, D ;
Ginty, DD ;
Vinson, C .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (02) :967-977
[3]   Expression of rat thyrotropin-releasing hormone (TRH) gene in TRH-producing tissues of transgenic mice requires sequences located in Exon 1 [J].
Balkan, W ;
Tavianini, MA ;
Gkonos, PJ ;
Roos, BA .
ENDOCRINOLOGY, 1998, 139 (01) :252-259
[4]  
BRADFORD J, 1976, ANAL BIOCHEM, V72, P248
[5]   Matlnspector and beyond: promoter analysis based on transcription factor binding sites [J].
Cartharius, K ;
Frech, K ;
Grote, K ;
Klocke, B ;
Haltmeier, M ;
Klingenhoff, A ;
Frisch, M ;
Bayerlein, M ;
Werner, T .
BIOINFORMATICS, 2005, 21 (13) :2933-2942
[6]   Cell-type-specific binding of the transcription factor CREB to the cAMP-response element [J].
Cha-Molstad, H ;
Keller, DM ;
Yochum, GS ;
Impey, S ;
Goodman, RH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (37) :13572-13577
[7]   Specificity proteins Sp1 and Sp3 interact with the rat GTP Cyclohydrolase I proximal promoter to regulate transcription [J].
Chandran, Nitya Sarath ;
Vunnavaj, Prashanthi ;
Wu, Yanning ;
Kapatos, Gregory .
JOURNAL OF NEUROCHEMISTRY, 2008, 104 (05) :1233-1248
[8]   HOMOLOGOUS CONDITIONED MEDIUM ENHANCES EXPRESSION OF TRH IN HYPOTHALAMIC NEURONS IN PRIMARY CULTURE [J].
CHARLI, JL ;
CRUZ, C ;
REDONDO, JL ;
GUERRA, C ;
JOSEPHBRAVO, P .
DEVELOPMENTAL BRAIN RESEARCH, 1995, 89 (01) :155-160
[9]   Minireview: Thyrotropin-Releasing Hormone and the Thyroid Hormone Feedback Mechanism [J].
Chiamolera, Maria Izabel ;
Wondisford, Fredric E. .
ENDOCRINOLOGY, 2009, 150 (03) :1091-1096
[10]   Dexamethasone represses cAMP rapid upregulation of TRH gene transcription:: identification of a composite glucocorticoid response element and a cAMP response element in TRH promoter [J].
Cote-Vélez, A ;
Pérez-Martínez, L ;
Díaz-Gallardo, MY ;
Pérez-Monter, C ;
Carreón-Rodríguez, A ;
Charli, JL ;
Joseph-Bravo, P .
JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2005, 34 (01) :177-197