Expression and function of variants of Slob, Slowpoke channel binding protein, in Drosophila
Expression and function of variants of Slob, Slowpoke channel binding protein, in Drosophila
复制标题
DOI：
10.1152/jn.00427.2005
复制
发表时间：
2006-03-01
影响因子：
2.5
通讯作者：
Levitan, IB
中科院分区：
文献类型：
--
作者：
Jaramillo, AM;Zeng, HY;Levitan, IB
Slob结合并调节果蝇Slowpoke（dSlo）钙激活钾通道，还将普遍存在的信号蛋白14 - 3 - 3募集到通道调节复合物中。RT-PCR揭示了果蝇头部存在多个slob转录本。这些转录本被预测编码我们称为Slob51（kDa），Slob57，Slob65和Slob71的蛋白质。Slob51和Slob65是缺少对14 - 3 - 3的结合重要的基序的剪接变体。以前的微阵列分析表明，昼夜节律循环的slob mRNA，我们通过定量PCR显示，在苍蝇头一个以上的转录周期。使用原位杂交，我们观察到不同的转录本的表达模式的差异。果蝇头部的免疫组织化学显示Slob71/65蛋白富集在外侧神经元中，与Slob57/51蛋白相反，Slob57/51蛋白在大脑间部神经元和背侧巨大中间神经元中表达最显著。使用异源表达系统，我们表明，不同的Slobs绑定到不同程度的dSlo和14 - 3 - 3。这些数据揭示了dSlo/Slob/14 - 3 - 3动态调控复合物的意外多样性。
Slob binds to and modulates the Drosophila Slowpoke ( dSlo) calcium-activated potassium channel and also recruits the ubiquitous signaling protein 14-3-3 to the channel regulatory complex. RT-PCR reveals the presence of multiple slob transcripts in Drosophila heads. The transcripts are predicted to encode proteins that we call Slob51 (kDa), Slob57, Slob65, and Slob71. Slob51 and Slob65 are splice variants that lack a motif important for the binding of 14-3-3. Previous microarray analyses demonstrated the circadian cycling of slob mRNA, and we show by quantitative PCR that more than one transcript cycles in fly heads. Using in situ hybridization, we observe differences in the expression patterns of the different transcripts. Immunohistochemistry on Drosophila heads reveals Slob71/65 protein to be enriched in the lateral neurons, in contrast to Slob57/51 protein, which is expressed most prominently in the pars intercerebralis neurons and dorsal giant interneurons. Using a heterologous expression system, we show that different Slobs bind to different extents to dSlo and 14-3-3. These data reveal an unexpected diversity of the dSlo/Slob/14-3-3 dynamic regulatory complex.