Differentiation-responsive elements in the 5' region of the mouse tissue plasminogen activator gene confer two-stage regulation by retinoic acid and cyclic AMP in teratocarcinoma cells.

Abstract

F9 cells induced to differentiate with retinoic acid (RA) increase transcription of the tissue plasminogen activator (t-PA) gene. Further treatment of these cells with cyclic AMP (cAMP) results in an additional stimulation of t-PA gene transcription. To investigate the mechanism of this two-stage regulation, 4 kilobase pairs (kbp) of 5'-flanking sequence from the murine t-PA gene was isolated. Two major start sites for transcription were found, neither of which depended on a classical TATA motif for correct initiation. By using transient transfection assays, it was determined that 4-kbp of flanking sequence could confer on reporter genes the same two-stage differentiation-specific expression as was observed for the endogenous t-PA gene. Deletion analyses of this 4-kbp fragment showed that 190 bp of flanking sequence was sufficient to bestow the same degree of two-stage regulation on reporter gene constructs. Within this region of DNA, sequence analysis revealed a possible cAMP regulatory element, a CTF/NF-1 recognition sequence, two potential Spl sites, and five potential binding sites for transcription factor AP-2. The deletion experiments, coupled with the positions of these potential cis-acting elements, suggest that multiple transcription factors, including those that bind to cAMP regulatory element, CTF/NF-1, Spl, and AP-2 sites, may be involved in regulation of the t-PA gene during F9 cell differentiation.

Transcriptional Regulation of the Transforming Growth Factor-β2 Promoter by cAMP-responsive Element-binding Protein (CREB) and Activating Transcription Factor-1 (ATF-1) Is Modulated by Protein Kinases and the Coactivators p300 and CREB-binding Protein

“…Finally, it is noteworthy that genes such as c-jun (87), FGF-3 (103), tissue plasminogen activator (104), and the TGF-␤ type II receptor (105) also contain CRE sites and turn on with RA-induced differentiation of EC cells. Like TGF-␤2, their promoters contain additional cis-regulatory elements that provide specificity and ensure that the correct gene is activated at the precise time.…”

Transcriptional Regulation of α1,3-Galactosyltransferase in Embryonal Carcinoma Cells by Retinoic Acid

“…ment of F9 Cells-To determine the temporal relationship between induction of the ␣1,3GT compared by RA to other known markers and to assess the efficiency of differentiation of F9 cells in response to 10 Ϫ7 M RA in our system, we determined the amount of laminin secreted into the cell culture media. It has been shown that the steady-state levels of mRNAs for laminin B1 and B2 increase dramatically following differentiation of F9 cells, and laminin has been defined as a differentiation marker, along with other basement membrane components, such as type IV collagen (␣1) (39,(55)(56)(57)(58)(59)(60). Laminin was detected by an enzyme-linked immunosorbent assay using cell culture media immobilized in microtiter wells and anti-laminin antibody as the detecting reagent.…”

GATA Factors and Androgen Receptor Collaborate To Transcriptionally Activate theRhox5Homeobox Gene in Sertoli Cells

“…This postmitotic somatic cell, which is in intimate contact with developing germ cells in seminiferous tubules in the testis, is crucial for all phases of male gametogenesis, including germ cell proliferation, meiosis, and differentiation (15). Genes necessary for the Sertoli cell to support male gametogenesis have been identified, and some progress has been made in identifying regulatory mechanisms that control the transcription of some of these genes, including follicle-stimulating hormone receptor (Fshr), cathepsin L, the inhibin-␤ B subunit, transferrin, Mullerian inhibitory substance, androgen-binding protein, Dnmrt 1, GATA1, GATA4, and tissue plasminogen activator (12,13,15,17,18,21,35,41,43,53). Regulatory regions and specific cis elements that participate in the transcriptional regulation of these genes have been identified in primary Sertoli cells and Sertoli cell lines.…”