Cyclic AMP inhibits fibronectin gene expression in a newly developed granulosa cell line by a mechanism that suppresses cAMP-responsive element-dependent transcriptional activation.
The main role of the ovarian granulosa cells is to nurse the oocyte and to produce estradiol and progesterone upon stimulation by gonadotropins. In fact, follicle-stimulating hormone (FSH) and luteinizing hormone control the expression of several genes during granulosa cell differentiation via cyclic AMP-dependent phosphorylations. Cyclic AMP stimulates transcription of genes that carry the cAMP-responsive element (CRE,5'TGACGTCA3') in their promoters. The fibronectin (FN) gene contains one CRE sequence at position -170. However, gonadotropins and cAMP inhibit FN gene expression in granulosa cells. To study the mechanism of the inhibition we developed a bovine granulosa cell line (BGC-1) that synthesizes estradiol in response to FSH and in which FSH and dibutyryl cAMP specifically decrease FN synthesis and its mRNA levels. The inhibitory effect (a) is not due to an alteration in FN mRNA stability, (b) requires upstream sequences other than CRE, located between positions -510 and -223, that are able to bind granulosa cell nuclear proteins, (c) is entirely dependent on the synthesis of intermediate proteins induced and or phosphorylated by cAMP, and (d) effectively suppresses the CRE-dependent transcriptional activation.
Bernath
V A
VA
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET), Buenos Aires, Argentina.
Muro
A F
AF
Vitullo
A D
AD
Bley
M A
MA
Barañao
J L
JL
Kornblihtt
A R
AR
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D003994:Bucladesine / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005353:Fibronectins / Q000235:genetics*; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006107:Granulosa Cells / Q000502:physiology*; D007621:Karyotyping; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D014158:Transcription, Genetic / Q000187:drug effects*
D004268:DNA-Binding Proteins; D005353:Fibronectins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D005576:Colforsin; D004958:Estradiol; D003994:Bucladesine; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP