Reference : Transcriptional induction of the human prolactin gene by cAMP requires two cis-acting...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/25927
Transcriptional induction of the human prolactin gene by cAMP requires two cis-acting elements and at least the pituitary-specific factor Pit-1
English
Peers, Bernard mailto [Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
Monget, Philippe [> > > >]
Nalda, M. Asuncion [> > > >]
Voz, Marianne mailto [Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
Berwaer, Monique [> > > >]
Belayew, Alexandra [> > > >]
Martial, Joseph mailto [Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
1991
Journal of Biological Chemistry
American Society for Biochemistry and Molecular Biology
266
27
18127-34
Yes (verified by ORBi)
International
0021-9258
1083-351X
Baltimore
MD
[en] Animals ; Base Sequence ; Binding Sites ; Blotting, Southern ; Blotting, Western ; Cattle ; Chloramphenicol O-Acetyltransferase/genetics ; Cyclic AMP/*metabolism ; DNA-Binding Proteins/*genetics/metabolism ; Humans ; Molecular Sequence Data ; Mutation ; Plasmids ; Prolactin/*genetics ; Promoter Regions, Genetic ; Rats ; Sequence Homology, Nucleic Acid ; Thymidine Kinase/genetics ; Transcription Factor Pit-1 ; Transcription Factors/*genetics/metabolism ; *Transcription, Genetic
[en] To identify the cis-acting elements responsible for cAMP stimulation of human prolactin (hPRL) promoter activity, pituitary GC cells were transfected with 5'-deleted hPRL promoters fused to the chloramphenicol acetyltransferase reporter gene. The proximal regulatory region (coordinates -250 to -42) was sufficient to confer strong cAMP stimulation (+/- 25 fold). Further 5' and 3' deletions performed within this proximal region demonstrated that two types of cis-acting elements are involved in the cAMP regulation: (i) the binding sites of the pituitary-specific factor Pit-1, and (ii) the sequence between coordinates -115 and -85 (named fragment A), which contains a TGACG motif. We show by gel-shift and Southwestern experiments that fragment A binds Pit-1 monomer and also a ubiquitous factor that is neither cAMP-responsive element-binding protein nor activator protein-1. Strong cAMP induction was observed when fragment A was juxtaposed to a Pit-1 binding site. That Pit-1 plays an important role was supported further by the finding that the hPRL proximal region conferred cAMP regulation when linked to the herpes simplex virus thymidine kinase promoter only in pituitary GC cells and not in other heterologous cells, which do not express Pit-1. Furthermore, we observed that concatenated Pit-1 binding sites were able to confer cAMP responsiveness to the thymidine kinase promoter in GC cells.
http://hdl.handle.net/2268/25927
http://www.jbc.org/cgi/reprint/266/27/18127?view=long&pmid=1655738
1991/09/25

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