"Shaping" of cell signaling via AKAP-tethered PDE4D: Probing with AKAR2-AKAP5 biosensor.

Q2 Biochemistry, Genetics and Molecular Biology Journal of Molecular Signaling Pub Date : 2012-05-14 DOI:10.1186/1750-2187-7-4

Salih S Koçer, Hsien-Yu Wang, Craig C Malbon

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引用次数: 10

Abstract

Unlabelled:

Background: PKA, a key regulator of cell signaling, phosphorylates a diverse and important array of target molecules and is spatially docked to members of the A-kinase Anchoring Protein (AKAP) family. AKAR2 is a biosensor which yields a FRET signal in vivo, when phosphorylated by PKA. AKAP5, a prominent member of the AKAP family, docks several signaling molecules including PKA, PDE4D, as well as GPCRs, and is obligate for the propagation of the activation of the mitogen-activated protein kinase cascade from GPCRs to ERK1,2.

Results: Using an AKAR2-AKAP5 fusion "biosensor", we investigated the spatial-temporal activation of AKAP5 undergoing phosphorylation by PKA in response to β-adrenergic stimulation. The pattern of PKA activation reported by AKAR2-AKAP5 is a more rapid and spatially distinct from those "sensed" by AKAR2-AKAP12. Spatial-temporal restriction of activated PKA by AKAP5 was found to "shape" the signaling response. Phosphatase PDE4D tethered to AKAP5 also later reverses within 60 s elevated intracellular cyclic AMP levels stimulated by β-adrenergic agonist. AKAP12, however, fails to attenuate the rise in cyclic AMP over this time. Fusion of the AKAP5 PDE4D-binding-domain to AKAP12 was found to accelerate a reversal of accumulation of intracellular cyclic AMP.

Conclusion: AKAPs, which are scaffolds with tethered enzymes, can "shape" the temporal and spatial aspects of cell signaling.

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通过AKAP-tethered PDE4D“塑造”细胞信号:用AKAR2-AKAP5生物传感器探测。

背景:PKA是细胞信号传导的关键调节因子，磷酸化多种重要的靶分子，并与a激酶锚定蛋白(AKAP)家族的成员在空间上对接。AKAR2是一种生物传感器，当被PKA磷酸化时，在体内产生FRET信号。AKAP5是AKAP家族的重要成员，对接包括PKA、PDE4D和GPCRs在内的几种信号分子，并专门用于从GPCRs到ERK1的丝裂原激活蛋白激酶级联的激活传播，2。结果:利用AKAR2-AKAP5融合“生物传感器”，我们研究了被PKA磷酸化的AKAP5在β-肾上腺素能刺激下的时空激活。与AKAR2-AKAP12“感知”的PKA激活模式相比，AKAR2-AKAP5报告的PKA激活模式更快速，在空间上也更明显。发现AKAP5对激活PKA的时空限制“塑造”了信号反应。与AKAP5连接的磷酸酶PDE4D随后也在60秒内逆转β-肾上腺素能激动剂刺激的细胞内环AMP水平升高。然而，在这段时间内，AKAP12不能减弱循环AMP的上升。研究发现，AKAP5 pde4d结合域与AKAP12的融合可加速逆转细胞内环amp的积累。结论:AKAPs作为拴系酶的支架，可以“塑造”细胞信号传导的时间和空间方面。

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Journal of Molecular Signaling Biochemistry, Genetics and Molecular Biology-Biochemistry

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期刊介绍： Journal of Molecular Signaling is an open access, peer-reviewed online journal that encompasses all aspects of molecular signaling. Molecular signaling is an exponentially growing field that encompasses different molecular aspects of cell signaling underlying normal and pathological conditions. Specifically, the research area of the journal is on the normal or aberrant molecular mechanisms involving receptors, G-proteins, kinases, phosphatases, and transcription factors in regulating cell proliferation, differentiation, apoptosis, and oncogenesis in mammalian cells. This area also covers the genetic and epigenetic changes that modulate the signaling properties of cells and the resultant physiological conditions.