AKAP13 Enhances CREB1 Activation by FSH in Granulosa Cells.
AKAP13 Enhances CREB1 Activation by FSH in Granulosa Cells.
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DOI：
10.1007/s43032-022-01097-5
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发表时间：
2023-05
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颗粒细胞（GCs）必须对促卵泡激素（FSH）有适当的反应，才能使卵泡成熟。FSH激活蛋白激酶A (PKA)，导致环AMP反应元件结合蛋白1 （CREB1）的磷酸化。我们鉴定了一种独特的a激酶锚定蛋白（AKAP13），该蛋白含有Rho鸟嘌呤核苷酸交换因子（RhoGEF）区域，在卵泡形成过程中诱导于GCs。已知akap可以协调信号级联反应，我们试图评估AKAP13在GCs对FSH的反应中的作用。过表达AKAP13的COV434人GCs中芳香酶报告活性升高。添加FSH或PKA激活剂forskolin可显著提高该活性，分别提高1.5- 2.5倍（p < 0.001）。PKA抑制剂H89显著降低了akap13依赖性的芳香化酶报告基因激活（p = 0.0067）。在共免疫沉淀实验中，AKAP13与CREB1发生物理相互作用，增加CREB1的磷酸化。FSH处理后CREB1磷酸化以特定时间的方式增加，而针对AKAP13的siRNA则降低了这种作用（p = 0.05）。在FSH存在的情况下，CREB1激活增加18.5倍，AKAP13共表达（p < 0.001）。芳香酶报告活性被RhoGEF区域、C3转移酶和A13抑制剂降低，而被RhoGEF激活剂A02大大增强。在原代小鼠和COV43 GCs中，与对照组相比，FSH处理细胞中Akap13/ Akap13的siRNA敲低降低了芳香化酶和黄体生成素受体转录。总的来说，这些发现表明AKAP13可能通过与CREB的相互作用在FSH信号转导中起支架蛋白的作用，导致CREB的磷酸化。
Granulosa cells (GCs) must respond appropriately to follicle-stimulating hormone (FSH) for proper follicle maturation. FSH activates protein kinase A (PKA) leading to phosphorylation of the cyclic AMP response element binding protein-1 (CREB1). We identified a unique A-kinase anchoring protein (AKAP13) containing a Rho guanine nucleotide exchange factor (RhoGEF) region that was induced in GCs during folliculogenesis. AKAPs are known to coordinate signaling cascades, and we sought to evaluate the role of AKAP13 in GCs in response to FSH. Aromatase reporter activity was increased in COV434 human GCs overexpressing AKAP13. Addition of FSH, or the PKA activator forskolin, significantly enhanced this activity by 1.5- to 2.5-fold, respectively (p < 0.001). Treatment with the PKA inhibitor H89 significantly reduced AKAP13-dependent activation of an aromatase reporter (p = 0.0067). AKAP13 physically interacted with CREB1 in co-immunoprecipitation experiments and increased the phosphorylation of CREB1. CREB1 phosphorylation increased after FSH treatment in a time-specific manner, and this effect was reduced by siRNA directed against AKAP13 (p = 0.05). CREB1 activation increased by 18.5-fold with co-expression of AKAP13 in the presence of FSH (p < 0.001). Aromatase reporter activity was reduced by inhibitors of the RhoGEF region, C3 transferase and A13, and greatly enhanced by the RhoGEF activator, A02. In primary murine and COV43 GCs, siRNA knockdown of Akap13/AKAP13 decreased aromatase and luteinizing hormone receptor transcripts in cells treated with FSH, compared with controls. Collectively, these findings suggest that AKAP13 may function as a scaffolding protein in FSH signal transduction via an interaction with CREB, resulting in phosphorylation of CREB.
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影响因子：
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作者：
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