Multiple binding sites for nuclear proteins of the anterior pituitary are located in the 5 '-flanking region of the porcine follicle-stimulating hormone (FSH) beta-subunit gene

Citation
Y. Kato et al., Multiple binding sites for nuclear proteins of the anterior pituitary are located in the 5 '-flanking region of the porcine follicle-stimulating hormone (FSH) beta-subunit gene, MOL C ENDOC, 158(1-2), 1999, pp. 69-78
Citations number
52
Categorie Soggetti
Endocrinology, Nutrition & Metabolism
Journal title
MOLECULAR AND CELLULAR ENDOCRINOLOGY
ISSN journal
03037207 → ACNP
Volume
158
Issue
1-2
Year of publication
1999
Pages
69 - 78
Database
ISI
SICI code
0303-7207(199912)158:1-2<69:MBSFNP>2.0.ZU;2-J
Abstract
Gonadotropins, follicle-stimulating hormone (FSH), and luteinizing hormone (LH), are synthesized specifically in the gonadotropes of the anterior pitu itary. The aim of this study was to investigate nuclear factors that bind s pecifically to the porcine FSH beta-subunit gene. We examined nuclear prote in binding to 2.75 kilobase pairs (kbp) of DNA adjacent to the porcine FSH beta-subunit gene: about 2.32 kbp of upstream DNA and 0.43 kbp of downstrea m DNA. The upstream region contains only TATA box, CACCC element, and some imperfect sequences of cAMP-responsive element, activator protein-1 binding site, and activator protein-2 binding site. Gel mobility shift assay using nuclear proteins;extracted from the porcine anterior pituitary revealed th at the proteins bound to a limited region of DNA, 107 bp long (designated a s Fd2), located about -800 bp upstream from the transcription initiation si te. Competitive binding assays demonstrated that the protein binding was se quence specific; the addition of excess amounts of several putative regulat ory sequences and plasmid (non-homologous) DNA fragments did not reduce the binding. Furthermore, all five subfragments of Fd2 were also bound: by the pituitary nuclear proteins, showing that the entire region of Fd2 is invol ved in this interaction. Southwestern blotting demonstrated that at least s even protein species of 110, 98, 78, 63, 52, 42, and 35 kDa recognize Fd2. Nuclear proteins from several other porcine tissues were also able to bind to the Fd2 fragment but the gel shift patterns were different and the bindi ngs were weak, although only the cerebellum showed a pattern of binding tha t was similar to that of the anterior pituitary. These data suggest that mu ltiple proteins of the anterior pituitary recognize a specific region of th e porcine FSH beta-subunit gene. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.