PKC-2 phosphorylation of UNC-18 Ser322 in AFD neurons regulates temperature dependency of locomotion.
PKC-2 phosphorylation of UNC-18 Ser322 in AFD neurons regulates temperature dependency of locomotion.
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DOI：
10.1523/jneurosci.4029-11.2012
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发表时间：
2012-05-16
期刊：
影响因子：
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通讯作者：
Barclay JW
中科院分区：
文献类型：
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作者：
Edwards MR;Johnson JR;Rankin K;Jenkins RE;Maguire C;Morgan A;Burgoyne RD;Barclay JW
二酰基甘油（DAG）/蛋白激酶C（PKC）信号传导在神经元功能的调节中起着不可或缺的作用。这在秀丽隐杆线虫中确实如此，特别是对于热敏信号和行为。然而，这种信号级联转导的下游分子靶标仍然几乎未被表征。我们研究了是否PKC磷酸化的Munc 18 -1，一个在囊泡运输和胞吐作用的必需蛋白，是下游效应DAG调节的热敏行为。我们在这里证明，C。Munc 18 -1的elegans直向同源物，Munc 18 -18，在体外在Ser 322处被磷酸化。用Ser 322磷酸突变体转基因拯救unc-18无效蠕虫显示出改变的温度敏感性。C. elegans表达3 DAG调节的PKC，阻断PKC-18 Ser 322磷酸化仅通过缺失钙激活的PKC-2进行表型模拟。非磷酸化的pkc-18 S322 A在AFD热敏神经元中的全神经元或特异性表达将野生型蠕虫转化为pkc-2无效表型。这些数据表明，一个单独的DAG依赖性的生物体的热敏行为的影响，特别是通过下游PKC-2磷酸化的丝氨酸322 AFD神经元上的PKC-18。
Diacylglycerol (DAG) / Protein Kinase C (PKC) signalling plays an integral role in the regulation of neuronal function. This is certainly true in Caenorhabditis elegans and in particular for thermosensory signalling and behaviour . Downstream molecular targets for transduction of this signalling cascade remain, however, virtually uncharacterised. We investigated whether PKC phosphorylation of Munc18-1, an essential protein in vesicle trafficking and exocytosis, was the downstream effector for DAG regulation of thermosensory behaviour. We demonstrate here that the C. elegans orthologue of Munc18-1, UNC-18, was phosphorylated in vitro at Ser322. Transgenic rescue of unc-18 null worms with Ser322 phosphomutants displayed altered thermosensitivity. C. elegans express 3 DAG-regulated PKCs and blocking UNC-18 Ser322 phosphorylation was phenocopied only by deletion of calcium-activated PKC-2. Expression of non-phosphorylatable UNC-18 S322A either panneuronally or specifically in AFD thermosensory neurons converted wildtype worms to a pkc-2 null phenotype. These data demonstrate that an individual DAG-dependent thermosensory behaviour of an organism is effected specifically by the downstream PKC-2 phosphorylation of UNC-18 on Ser322 in AFD neurons.