ANKS3 Co-Localises with ANKS6 in Mouse Renal Cilia and Is Associated with Vasopressin Signaling and Apoptosis In Vivo in Mice.
ANKS3 Co-Localises with ANKS6 in Mouse Renal Cilia and Is Associated with Vasopressin Signaling and Apoptosis In Vivo in Mice.
复制标题
ANKS3在小鼠肾纤毛中与ANKS6共定位，并与小鼠体内的加压素信号传导和凋亡相关。
DOI：
10.1371/journal.pone.0136781
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Gauguier D
中科院分区：
文献类型：
--
作者：
Delestré L;Bakey Z;Prado C;Hoffmann S;Bihoreau MT;Lelongt B;Gauguier D
Ankyrin重复序列和含有6个不育的α基序结构域(ANKS6)的突变在多囊肾病PKD/MHM(Cy/+)大鼠模型的肾囊变和人类肾小球肾炎中起重要作用。ANKS6的蛋白质伙伴网络正在形成，其功能特征为理解ANKS6在肾脏生物学中的作用和参与肾囊形成的机制提供了重要线索。在使用酵母双杂交系统实验证实ANKS6和ANKS3之间的相互作用之后，我们证明了这两种蛋白之间的结合是通过它们的无菌α基序(SAM)进行的，并且大鼠ANSK6中的823氨基酸是这种相互作用的关键。我们通过免疫共沉淀进一步显示了它们的相互作用，并在小鼠体内证明了ANKS3存在于肾纤毛中。锁定核酸(LNA)修饰的反义寡核苷酸下调了小鼠体内Anks3的表达，与血管加压素诱导基因的转录增加有关，这表明肾脏的水通透性发生了变化，并改变了与纤毛结构、细胞凋亡和细胞增殖相关的编码蛋白质的基因的转录。这些数据提供了ANKS3-ANKS6通过其SAM结构域和共定位在小鼠肾纤毛中直接相互作用的实验证据，并阐明了ANKS6间接介导的小鼠肾脏中可能受ANKS3-ANKS6相互作用改变影响的分子机制。我们的结果有助于更好地了解与ANKS6相互作用的蛋白质网络的结构和功能，这可能是囊性疾病的治疗靶点。
Mutations in Ankyrin repeat and sterile alpha motif domain containing 6 (ANKS6) play a causative role in renal cyst formation in the PKD/Mhm(cy/+) rat model of polycystic kidney disease and in nephronophthisis in humans. A network of protein partners of ANKS6 is emerging and their functional characterization provides important clues to understand the role of ANKS6 in renal biology and in mechanisms involved in the formation of renal cysts. Following experimental confirmation of interaction between ANKS6and ANKS3 using a Yeast two hybrid system, we demonstrated that binding between the two proteins occurs through their sterile alpha motif (SAM) and that the amino acid 823 in rat ANSK6 is key for this interaction. We further showed their interaction by co-immunoprecipitation and showed in vivo in mice that ANKS3 is present in renal cilia. Downregulated expression of Anks3 in vivo in mice by Locked Nucleic Acid (LNA) modified antisense oligonucleotides was associated with increased transcription of vasopressin-induced genes, suggesting changes in renal water permeability, and altered transcription of genes encoding proteins involved in cilium structure, apoptosis and cell proliferation. These data provide experimental evidence of ANKS3-ANKS6 direct interaction through their SAM domain and co-localisation in mouse renal cilia, and shed light on molecular mechanisms indirectly mediated by ANKS6 in the mouse kidney, that may be affected by altered ANKS3-ANKS6 interaction. Our results contribute to improved knowledge of the structure and function of the network of proteins interacting with ANKS6, which may represent therapeutic targets in cystic diseases.