Exogenous expression of Pit-1 in AtT-20 corticotropic cells induces endogenous growth hormone gene transcription.
The pituitary-specific POU-homeodomain transcription factor, Pit-1, is known to regulate the expression of the GH gene in somatotropes, prolactin (PRL) in lactotropes, and TSH in thyrotropes. It is not normally expressed in corticotropes or gonadotropes. We addressed the question of whether exogenous Pit-1 was sufficient to induce ectopic transcription of the GH gene in the corticotropic cell line, AtT-20, or the gonadotropic cell line, alpha T3-1. A fusion gene composed of enhanced green fluorescent protein gene and human Pit-1 cDNA was transfected into AtT-20 and alpha T3-1 cells. The endogenous mouse GH mRNA was induced in three of nine AtT-20 cell lines and one of three alpha T3-1 cell lines containing the fusion gene. A small amount of GH protein was also detected in these cell lines. These data indicate that transfected Pit-1 is capable of inducing transcription of the GH gene in AtT-20 cells and alpha T3-1 cells. These data also suggest that synergistic co-factors might be required to transcribe the GH gene effectively for translation into GH protein. Furthermore, our findings support the hypothesis that the function of anterior pituitary cells is determined by the combinatorial action of specific transcription factors.
Kurotani
R
R
Department of Pathology, Tokai University School of Medicine, Boseidai, Isehara, Kanagawa 259-1193, Japan.
Yoshimura
S
S
Iwasaki
Y
Y
Inoue
K
K
Teramoto
A
A
Osamura
R Y
RY
D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D013006:Growth Hormone / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008815:Mice, Inbred Strains; D018613:Microscopy, Confocal; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050817:Transcription Factor Pit-1; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000324:Adrenocorticotropic Hormone; D004268:DNA-Binding Proteins; D049452:Green Fluorescent Proteins; D013006:Growth Hormone; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D050817:Transcription Factor Pit-1; D014157:Transcription Factors; C494187:POU1F1 protein, human; C494185:Pou1f1 protein, mouse