Document Detail


Two proximal activating protein-1-binding sites are sufficient to stimulate transcription of the ovine follicle-stimulating hormone-beta gene.
MedLine Citation:
PMID:  9165057     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
FSH is an important regulator of mammalian gametogenesis and the female reproductive cycle. Although little is known about the transcriptional regulation of the beta-subunit (the rate-limiting subunit of FSH synthesis), sequence analysis of the ovine FSHbeta promoter has revealed a number of potential activating protein-1 (AP-1; Jun/Fos)-binding sites. To determine whether the gene encoding the beta-subunit of ovine FSH (oFSHbeta) is responsive to AP-1 transcriptional complexes, chimeric constructs containing deleted portions of the oFSHbeta promoter fused to a luciferase reporter were transiently transfected along with c-Jun and c-Fos expression constructs into JAR cells. Analysis of these deletion constructs revealed that the proximal promoter of oFSHbeta is highly stimulated by c-Jun and c-Fos proteins (typically 20-fold with a reporter construct containing oFSHbeta sequences from -215 to +759 bp). This stimulation was lost when a similar construct containing sequences from -84 to +759 bp was tested. Transcriptional start site analysis using reverse transcription-PCR verified that the transcriptional initiation of the -215-bp deletion construct, with or without cotransfected c-Jun/c-Fos, was the same as that observed in vivo. Computer analysis of oFSHbeta sequences from -215 to +1 bp identified four putative AP-1-like elements, located at -155, -120, -83, and -10 bp. Gel retardation experiments using oligonucleotides corresponding to the four putative AP-1-like sites revealed that only -120 and -83 sites in oFSHbeta bound AP-1 proteins in vitro. Site-directed mutagenesis of the -120 and -83 sites showed that each element was required for stimulation by c-Jun and c-Fos proteins as well as 12-O-tetradecanoyl phorbol-13-acetate in transient transfection assays. Finally, immunocytochemical dual labeling was used to show that more than 75% of all FSHbeta-containing cells in ovine pituitary sections from cycling ewes contained nuclear c-Jun, JunB, JunD, and Fos proteins. These data, taken together, show that oFSHbeta transcription can be stimulated by c-Jun and c-Fos proteins via two functionally linked AP-1-like sites in the oFSHbeta proximal promoter and that these sites are likely to be important regulators of FSH production in vivo.
Authors:
B D Strahl; H J Huang; N R Pedersen; J C Wu; B R Ghosh; W L Miller
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Endocrinology     Volume:  138     ISSN:  0013-7227     ISO Abbreviation:  Endocrinology     Publication Date:  1997 Jun 
Date Detail:
Created Date:  1997-06-17     Completed Date:  1997-06-17     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0375040     Medline TA:  Endocrinology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2621-31     Citation Subset:  AIM; IM    
Affiliation:
Department of Biochemistry, North Carolina State University, Raleigh 27695-7622, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Binding Sites
Cattle
Choriocarcinoma
Consensus Sequence
DNA Primers
Female
Follicle Stimulating Hormone / biosynthesis*,  genetics*
Follicle Stimulating Hormone, beta Subunit
Globins / biosynthesis
Hela Cells
Humans
Luciferases / biosynthesis
Molecular Sequence Data
Polymerase Chain Reaction
Proto-Oncogene Proteins c-fos / biosynthesis
Proto-Oncogene Proteins c-jun / biosynthesis
Rabbits
Rats
Recombinant Fusion Proteins / biosynthesis
Sequence Deletion
Sheep
Swine
Transcription Factor AP-1 / metabolism*
Transcription, Genetic*
Transfection
Tumor Cells, Cultured
Uterine Neoplasms
Grant Support
ID/Acronym/Agency:
NICHD 10773/HD/NICHD NIH HHS; NICHD 34863/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
0/DNA Primers; 0/Follicle Stimulating Hormone, beta Subunit; 0/Proto-Oncogene Proteins c-fos; 0/Proto-Oncogene Proteins c-jun; 0/Recombinant Fusion Proteins; 0/Transcription Factor AP-1; 9002-68-0/Follicle Stimulating Hormone; 9004-22-2/Globins; EC 1.13.12.-/Luciferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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