Gravin dynamics regulates the subcellular distribution of PKA.
Gravin dynamics regulates the subcellular distribution of PKA.
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DOI：
10.1016/j.yexcr.2008.12.026
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发表时间：
2009-04-15
影响因子：
3.7
通讯作者：
Grove B
中科院分区：
文献类型：
--
作者：
Yan X;Walkiewicz M;Carlson J;Leiphon L;Grove B
Gravin是一种多价A激酶锚定蛋白（AKAP），定位于几种细胞类型的细胞外周，并被认为将PKA和其他结合伴侣靶向质膜。一个N-末端豆蔻酰化序列和三个区域丰富的碱性氨基酸被提出来介导这种定位。有报道表明佛波醇酯影响Gravin的大鼠直系同源物SSeCKS的分布，进一步表明PKC也可能调节Gravin的亚细胞分布，这反过来又可能影响PKA的分布。在这项研究中，表达全长和突变体gravin-EGFP构建体缺乏拟议的靶向结构域的细胞的定量共聚焦显微镜显示，无论是N-豆蔻酰化位点或多碱性区域足以将gravin靶向细胞外周。此外，佛波酯处理诱导gravin-EGFP从细胞外周到胞核囊泡隔室的再分布，但这需要N-肉豆蔻酰化位点的存在。共聚焦显微镜进一步揭示，不仅gravin-EGFP将PKA RII-ECFP构建体靶向细胞外周，而且PKC激活导致gravin和PKA构建体重新分布到相同的亚细胞位点。推测gravin对PKC活性的这种动态响应可能介导PKC依赖性的PKA活性控制。
Gravin, a multivalent A-kinase anchoring protein (AKAP), localizes to the cell periphery in several cell types and is postulated to target PKA and other binding partners to the plasma membrane. An N-terminal myristoylation sequence and three regions rich in basic amino acids are proposed to mediate this localization. Reports indicating that phorbol ester affects the distribution of SSeCKS, the rat orthologue of gravin, further suggest that PKC may also regulate the subcellular distribution of gravin, which in turn may affect PKA distribution. In this study, quantitative confocal microscopy of cells expressing full-length and mutant gravin-EGFP constructs lacking the proposed targeting domains revealed that either the N-myristoylation site or the polybasic regions were sufficient to target gravin to the cell periphery. Moreover, phorbol ester treatment induced redistribution of gravin-EGFP from the cell periphery to a juxtanuclear vesicular compartment, but this required the presence of the N-myristoylation site. Confocal microscopy further revealed that not only did gravin-EGFP target a PKA RII-ECFP construct to the cell periphery, but PKC activation resulted in redistribution of the gravin and PKA constructs to the same subcellular site. It is postulated that this dynamic response by gravin to PKC activity may mediate PKC dependent control of PKA activity.
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影响因子：
1.7
作者：
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通讯作者：
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