title: The proximal Gata4 promoter directs reporter gene expression to sertoli cells during mouse gonadal development.
abstract: The GATA4 transcription factor is an important developmental determinant for many organs, such as the heart, gut, and testis. Despite this pivotal role, our understanding of the transcriptional mechanisms that control the proper spatiotemporal expression of the GATA4 gene remains limited. We have generated transgenic mice expressing a green fluorescent protein (GFP) marker under the control of rat Gata4 5' flanking sequences. Several GATA4-expressing organs displayed GFP fluorescence, including the heart, intestine, and pancreas. In the gonads, while GATA4 is expressed in pregranulosa, granulosa, and theca ovarian cells, and Sertoli, Leydig, and peritubular testicular cells, the first 5 kb of Gata4 regulatory sequences immediately upstream of exon 1 were sufficient to direct GFP reporter expression only in testis and, specifically, in Sertoli cells. Onset of GFP expression occurred after Sertoli cell commitment and was maintained in these cells throughout development to adulthood. In vitro studies revealed that the first 118 bp of the Gata4 promoter is sufficient for full basal activity in several GATA4-expressing cell lines. Promoter mutagenesis and DNA-binding experiments identified two GC-box motifs and, particularly, one E-box element within this -118-bp region that are crucial for its activity. Further analysis revealed that members of the USF family of transcription factors, especially USF2, bind to and activate the Gata4 promoter via this critical E-box motif.
authors.affiliation: Ontogeny-Reproduction Research Unit, Centre de Recherche du Centre Hospitalier Universitaire de Québec, Centre de Recherche en Biologie de la Reproduction, Department of Obstetrics and Gynecology, Laval University, Québec City, Québec, Canada G1K 7P4.
authors.forename: Séverine
authors.initials: S
authors.lastname: Mazaud Guittot
authors.forename: Amélie
authors.initials: A
authors.lastname: Tétu
authors.forename: Eric
authors.initials: E
authors.lastname: Legault
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pilon
authors.forename: David W
authors.initials: DW
authors.lastname: Silversides
authors.forename: Robert S
authors.initials: RS
authors.lastname: Viger
mesh_terms: D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D024721:E-Box Elements / Q000235:genetics; D050991:GATA4 Transcription Factor / Q000032:analysis / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012708:Sertoli Cells / Q000737:chemistry / Q000378:metabolism*; D012727:Sex Characteristics; D013737:Testis / Q000166:cytology / Q000196:embryology* / Q000254:growth & development*; D019076:Transgenes
chemical_list: D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; D049452:Green Fluorescent Proteins