Regulatory elements controlling pituitary-specific expression of the human prolactin gene.
We have performed transfection and DNase I footprinting experiments to investigate pituitary-specific expression of the human prolactin (hPRL) gene. When fused to the chloramphenicol acetyltransferase (CAT) reporter gene, 5,000 base pairs of the 5'-flanking sequences of the hPRL gene were able to drive high cat gene expression in prolactin-expressing GH3B6 cells specifically. Deletion analysis indicated that this pituitary-specific expression was controlled by three main positive regulatory regions. The first was located just upstream from the TATA box between coordinates -40 and -250 (proximal region). We have previously shown that three motifs of this region bind the pituitary-specific Pit-1 factor. The second positive region was located in the vicinity of coordinates -1300 to -1750 (distal region). DNase I footprinting assays revealed that eight DNA motifs of this distal region bound protein Pit-1 and that two other motifs were recognized by ubiquitous factors, one of which seems to belong to the AP-1 (jun) family. The third positive region was located further upstream, between -3500 and -5000 (superdistal region). This region appears to enhance transcription only in the presence of the distal region.
Peers
B
B
Université de Liège, Institut de Chimie B6, Sart-Tilman, Belgium.
Voz
M L
ML
Monget
P
P
Mathy-Hartert
M
M
Berwaer
M
M
Belayew
A
A
Martial
J A
JA
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003850:Deoxyribonuclease I; D005796:Genes; D005809:Genes, Regulator; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010911:Pituitary Neoplasms; D011388:Prolactin / Q000235:genetics*; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D015345:Oligonucleotide Probes; D011388:Prolactin; D015500:Chloramphenicol O-Acetyltransferase; D003850:Deoxyribonuclease I