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Article (Scientific journals)
Pit-1 binding sequences permit calcium regulation of human prolactin gene expression
Hoggard, Nigel; Davis, Julian R E; Berwaer, Monique et al.
1991In Molecular Endocrinology, 5 (11), p. 1748-54
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Keywords :
Aminoquinolines; Animals; Base Sequence; Binding Sites; Calcium/*physiology; Chloramphenicol O-Acetyltransferase/genetics/metabolism; Chromosome Deletion; DNA-Binding Proteins/*metabolism; Fluorescent Dyes; Genetic Vectors; Humans; Molecular Sequence Data; Oligodeoxyribonucleotides; Pituitary Neoplasms; Prolactin/*genetics; *Promoter Regions, Genetic; Rats; Recombinant Fusion Proteins/metabolism; Spectrometry, Fluorescence; Thymidine Kinase/genetics/metabolism; Transcription Factor Pit-1; Transcription Factors/*metabolism; *Transfection
Abstract :
[en] This study examines the regulation of the human PRL (hPRL) gene promoter by intracellular calcium. Deletants of the 5'-flanking region of the hPRL gene and constructs consisting of the thymidine kinase promoter linked to the first or second proximal Pit-1 binding site were fused to the bacterial chloramphenicol acetyl transferase (CAT) reporter gene. With the complete 5-kilobase pair (kbp) hPRL promoter sequence the calcium channel agonist Bay K8644 induced a significant 2-fold increase in CAT reporter gene expression and the antagonist verapamil a 4.5-fold reduction, using GH3 cells cultured in physiological levels of calcium. The transcriptional response to calcium influx was similar with a series of 5'-deleted hPRL-CAT constructs including those that comprised the proximal (up to 740 bp) or distal (-1300- to -1700-bp) sequences alone. When treating cells cultured in low calcium conditions the induction with the hPRL promoter increased to 5-fold on the addition of exogenous calcium and Bay K8644. The pituitary-specific expression of the hPRL gene is conferred by the interaction of the pituitary-specific factor Pit-1 with several binding sites located in the 5'-flanking DNA, of which three are located in the proximal region. This suggested that Pit-1 binding sites may be involved in the calcium response.(ABSTRACT TRUNCATED AT 250 WORDS)
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Hoggard, Nigel
Davis, Julian R E
Berwaer, Monique
Monget, Philippe
Peers, Bernard ;  Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Belayew, Alexandra
Martial, Joseph ;  Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Language :
English
Title :
Pit-1 binding sequences permit calcium regulation of human prolactin gene expression
Publication date :
1991
Journal title :
Molecular Endocrinology
ISSN :
0888-8809
eISSN :
1944-9917
Publisher :
Endocrine Society, Chevy Chase, United States - Maryland
Volume :
5
Issue :
11
Pages :
1748-54
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 27 August 2009

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