Structure of the rat inhibin and activin βA-subunit gene and regulation in an ovarian granulosa cell line

被引:35
作者
Ardekani, AM [1 ]
Romanelli, JCD [1 ]
Mayo, KE [1 ]
机构
[1] Northwestern Univ, Dept Biochem Mol Biol & Cell Biol, Evanston, IL 60208 USA
关键词
D O I
10.1210/en.139.7.3271
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We have isolated the rat inhibin and activin beta(A)-subunit gene, which is composed of three exons, and have characterized a 571-bp region upstream from the transcriptional start site that functions as a promoter in transient transfection studies in an ovarian granulosa cell line, GRMO2. Deletion analysis of the 571-bp promoter region has identified DNA sequences between -362 bp and -110 bp to be essential in mediating basal promoter activity and activation by forskolin (FSK) and/or 12-O-tetradecanoylphorbol-13-acetate (TPA). Within this region, a variant CRE (cAMP response element) has been identified at -120 bp. Point mutations in the variant CRE substantially reduce the ability of FSK and/or TPA to induce promoter activity in GRMO2 cells. A single nucleotide change in the variant CRE, which converts it to a consensus CRE, does not enhance promoter activity in response to FSK and/or TPA, but rather reduces promoter activity to the same extent as the other inactivating mutation in the variant CRE, suggesting that this element does not act as a classical CRE. Consistent with this, electrophoretic mobility shift assays performed using antibodies to a variety of cAMP and phorbol ester-responsive transcription factors indicate that the AP-1 family proteins jun-B and fos-B are present in the protein complex binding to the variant CRE. Overexpression of jun-B and fos-B in GRMO2 cells resulted in a robust activation of the beta(A)-subunit promoter. Our results suggest that this novel variant CRE sequence mediates both cAMP and phorbol ester regulation through its interactions with AP-1 family proteins.
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页码:3271 / 3279
页数:9
相关论文
共 52 条
[1]   EXPRESSION OF THE GLYCOPROTEIN HORMONE ALPHA-SUBUNIT GENE IN THE PLACENTA REQUIRES A FUNCTIONAL CYCLIC-AMP RESPONSE ELEMENT, WHEREAS A DIFFERENT CIS-ACTING ELEMENT MEDIATES PITUITARY-SPECIFIC EXPRESSION [J].
BOKAR, JA ;
KERI, RA ;
FARMERIE, TA ;
FENSTERMAKER, RA ;
ANDERSEN, B ;
HAMERNIK, DL ;
YUN, J ;
WAGNER, T ;
NILSON, JH .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (11) :5113-5122
[2]   CHARACTERIZATION OF IMMORTALIZED MOUSE GRANULOSA-CELL LINES [J].
BRIERS, TW ;
VANDEVOORDE, A ;
VANDERSTICHELE, H .
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 1993, 29A (11) :847-854
[3]  
BRUZDZINSKI CJ, 1990, J BIOL CHEM, V265, P2078
[4]  
CAMPBELL MJ, 1995, BIOTECHNIQUES, V18, P1027
[5]   HORMONAL-CONTROL OF GENE-EXPRESSION - MULTIPLICITY AND VERSATILITY OF CYCLIC ADENOSINE 3',5'-MONOPHOSPHATE-RESPONSIVE NUCLEAR REGULATORS [J].
DEGROOT, RP ;
SASSONECORSI, P .
MOLECULAR ENDOCRINOLOGY, 1993, 7 (02) :145-153
[6]  
DEUTSCH PJ, 1988, J BIOL CHEM, V263, P18466
[7]   FIREFLY LUCIFERASE GENE - STRUCTURE AND EXPRESSION IN MAMMALIAN-CELLS [J].
DEWET, JR ;
WOOD, KV ;
DELUCA, M ;
HELINSKI, DR ;
SUBRAMANI, S .
MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (02) :725-737
[8]   BINDING-SPECIFICITY OF CYCLIC ADENOSINE 3',5'-MONOPHOSPHATE-RESPONSIVE ELEMENT (CRE)-BINDING PROTEINS AND ACTIVATING TRANSCRIPTION FACTORS TO NATURALLY-OCCURRING CRE SEQUENCE VARIANTS [J].
DRUST, DS ;
TROCCOLI, NM ;
JAMESON, JL .
MOLECULAR ENDOCRINOLOGY, 1991, 5 (10) :1541-1551
[9]   2 MESSENGER RIBONUCLEIC-ACIDS ENCODING THE COMMON BETA(B)-CHAIN OF INHIBIN AND ACTIVIN HAVE DISTINCT 5'-INITIATION SITES AND ARE DIFFERENTIALLY REGULATED IN RAT GRANULOSA-CELLS [J].
DYKEMA, JC ;
MAYO, KE .
ENDOCRINOLOGY, 1994, 135 (02) :702-711
[10]  
DYKEMA JC, 1993, 75 ANN M END SOC LAS, P1252