Follicle-stimulating Hormone Stimulates Protein Kinase A-mediated Histone H3 Phosphorylation and Acetylation Leading to Select Gene Activation in Ovarian Granulosa Cells

Abstract

We examined the phosphorylation and acetylation of histone H3 in ovarian granulosa cells stimulated to differentiate by follicle-stimulating hormone (FSH). We found that protein kinase A (PKA) mediates H3 phosphorylation on serine 10, based on inhibition exclusively by PKA inhibitors. FSH-stimulated H3 phosphorylation in granulosa cells is not downstream of mitogenactivated protein kinase/extracellular signal-regulated kinase, ribosomal S6 kinase-2, mitogen-and stressactivated protein kinase-1, p38 MAPK, phosphatidylinositol-3 kinase, or protein kinase C. Transcriptional activation-associated H3 phosphorylation on serine 10 and acetylation of lysine 14 leads to activation of serum glucocorticoid kinase, inhibin ␣, and c-fos genes. We propose that phosphorylation of histone H3 on serine 10 by PKA in coordination with acetylation of H3 on lysine 14 results in reorganization of the promoters of select FSH responsive genes into a more accessible configuration for activation. The unique role of PKA as the physiological histone H3 kinase is consistent with the central role of PKA in initiating granulosa cell differentiation.Maturation of ovarian follicles to a preovulatory stage requires follicle-stimulating hormone (FSH) 1 production by the pituitary gland. The FSH receptor is a member of the G protein-coupled seven-transmembrane receptor family and is coupled to adenylyl cyclase (1). It is expressed exclusively on ovarian granulosa cells in female mammals (2). Most of the actions of FSH are mediated by cAMP formation and activation of protein kinase A (PKA), based on the ability of cell-permeable cAMP analogs to mimic the known differentiation responses to FSH in granulosa cells (2) and on the ability of the PKA inhibitors H89 2 (3) and KT5720 2 to inhibit granulosa cell differentiation. The downstream consequences of FSH are well established and include, for example, the induction of receptors for luteinizing hormone (LH) and prolactin, induction of enzymes associated with the increased steroidogenic capacity of granulosa cells including P450 aromatase and cholesterol side chain cleavage, induction of proteins associated with PKA signaling including RII␤ (2, 4) and AKAP80 (5), and expression of the hormone inhibin (6). However, gene and/or protein induction for these responses to FSH is generally delayed by at least 24 h (2-4, 7). The more immediate responses to FSH, which lead to the induction of immediate early genes such as c-fos and serum glucocorticoid kinase (SGK) (3,8), are less well understood. Further elucidation of the FSH signaling pathways that lead to the induction of immediate early genes would be useful to understand how FSH initiates granulosa cell differentiation. We have previously shown that FSH (via PKA) promotes activation of the p42/p44 mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway (9). FSH also activates the p38 MAPK pathway (10) and downstream phosphorylation of the heat shock protein (HSP) 27, leading to granulosa cell rounding (10). We 3 and ot...

cAMP signaling induces rapid loss of histone H3 phosphorylation in mammary adenocarcinoma-derived cell lines

“…Our observation of rapid, cAMP-mediated loss of H3 phosphorylation stands in contrast to that of Salvador and colleagues who reported that cAMP signaling rapidly and transiently induces H3 phosphorylation in primary cultures of ovarian granulosa cells and at cAMP target promoters [11]. Clearly, the response of a given cell to cAMP signaling depends on the array of cyclic nucleotide binding proteins and downstream effectors expressed.…”

“…An interesting aspect of H3 phosphorylation is its regulation by signal transduction pathways. Phosphorylation of histone H3 at serine 10 has been shown to be increased in mammalian interphase nuclei by the MAP kinase [9], protein kinase A (PKA) [10,11], and NF-κB-cytokine signaling pathways [12,13]. In each case increased H3 phosphorylation was found to occur at particular genes activated by these pathways which suggests a role in regulation of gene expression [11][12][13][14][15].…”

“…Phosphorylation of histone H3 at serine 10 has been shown to be increased in mammalian interphase nuclei by the MAP kinase [9], protein kinase A (PKA) [10,11], and NF-κB-cytokine signaling pathways [12,13]. In each case increased H3 phosphorylation was found to occur at particular genes activated by these pathways which suggests a role in regulation of gene expression [11][12][13][14][15]. Kinases shown to play important roles in the regulation of H3 phosphorylation at serines 10 and 28 include MSK1/2 [16,17], IKKα [12,13] and aurora kinases (mitotic phosphorylation) [18].…”

A phosphatase cascade by which rewarding stimuli control nucleosomal response

“…We therefore investigated the phosphorylation of histone H3 on Ser-10, a substrate for several kinases 28,29 , which is known to be regulated by PP1 30,31 . The phosphorylation of H3 on Ser-10 is a key step in nucleosomal response 28,32 and is crucial for memory formation 33 .…”

Phosphorylation at serine 28 and acetylation at lysine 9 of histone H3 induced by trichostatin A

“…Besides acetylation of histones, the core histones and histone H1 undergo phosphorylation on specific serine and threonine residues in chromosome condensation during mitosis (Bradbury, 1992;Koshland and Strunnikov, 1996). Phosphorylation of histone H3 N-terminal serine 10 is closely related to both the induction of immediate-early (IE) genes, including proto-oncogenes c-fos and c-jun, and to chromatin remodeling and chromosome condensation during mitosis and meiosis (Wei et al, 1998;Chadee et al, 1999;Sassone-Corsi et al, 1999;Thomson et al, 1999;Clayton et al, 2000;Salvador et al, 2001). In addition, a recent study has shown that phosphorylation of histone H3 at serine 28 also occurs in chromosomes predominantly during early mitosis and coincides with the initiation of mitotic chromosome condensation in various mammalian cell lines (Goto et al, 1999).…”