Chromodomain helicase binding protein 8 (Chd8) is a novel A-kinase anchoring protein expressed during rat cardiac development.

Chromodomain helicase binding protein 8 (Chd8) is a novel A-kinase anchoring protein expressed during rat cardiac development.
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DOI:
10.1371/journal.pone.0046316
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Bond M
Bond M
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Shanks MO;Lund LM;Manni S;Russell M;Mauban JR;Bond M

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A-激酶锚定蛋白(AKAP)结合蛋白激酶A(PKA)的调节亚基,并将全酶定位于多个亚细胞隔间中离散的信号微域。尽管有新的证据表明,PKA核池对PKA信号级联信号通路的激活迅速做出反应,但仅有少数AKAP定位于细胞核。在这里,我们显示了CHD8的氨基末端有一个PKA结合域,并证明了CHD8与RII的亚细胞共存。RII重叠和免疫沉淀实验表明CHD8-S与RIIα结合。一旦RIIα去磷酸化,结合就被取消。通过免疫荧光,我们在HeLa细胞和新生大鼠心肌细胞中发现了CHD8的核和核周池。我们还发现,从出生后的大鼠心脏提取的RNA中有高水平的CHD8 mRNA。这些数据将CHD8添加到已知的核AKAP的短名单中,并暗示CHD8在出生后大鼠心脏发育中的功能。
A-kinase anchoring proteins (AKAPs) bind the regulatory subunits of protein kinase A (PKA) and localize the holoenzyme to discrete signaling microdomains in multiple subcellular compartments. Despite emerging evidence for a nuclear pool of PKA that rapidly responds to activation of the PKA signaling cascade, only a few AKAPs have been identified that localize to the nucleus. Here we show a PKA-binding domain in the amino terminus of Chd8, and demonstrate subcellular colocalization of Chd8 with RII. RII overlay and immunoprecipitation assays demonstrate binding between Chd8-S and RIIα. Binding is abrogated upon dephosphorylation of RIIα. By immunofluorescence, we identified nuclear and perinuclear pools of Chd8 in HeLa cells and rat neonatal cardiomyocytes. We also show high levels of Chd8 mRNA in RNA extracted from post-natal rat hearts. These data add Chd8 to the short list of known nuclear AKAPs, and implicate a function for Chd8 in post-natal rat cardiac development.
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