Identification of RNA aptamer which specifically interacts with PtdIns(3)P
Identification of RNA aptamer which specifically interacts with PtdIns(3)P
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DOI：
10.1016/j.bbrc.2019.07.034
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发表时间：
2019-09-10
影响因子：
3.1
通讯作者：
Noguchi, Masayuki
中科院分区：
文献类型：
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作者：
Donia, Thoria;Jyoti, Bala;Noguchi, Masayuki
磷脂酰肌醇Ptdlns(3)P在自噬中起重要作用，但其活性的具体机制尚不清楚。在这里，我们使用指数浓缩配体的系统进化(SELEX)筛选方法来鉴定一个40个核苷酸的RNA适配子，它特异性地识别并结合细胞内溶酶体Ptdlns(3)P。结合以一种依赖于镁浓度和pH的方式发生，从而抑制自噬，如通过LC 3 II/I转换、P62降解、LC 3斑点的形成和Phafin2的溶酶体积累所确定的。这些作用反过来又抑制了溶酶体的酸化，以及自噬诱导后组织蛋白酶D随后的水解性。鉴于Ptdlns(3)P作为诱导自噬的关键靶向分子的重要作用，这一新型Ptdlns(3)P RNA适配子的鉴定为研究磷脂酰肌醇的生物学功能和机制提供了新的机会。(C)2019年提交人。由爱思唯尔公司出版。
The phosphinositide Ptdlns(3)P plays an important role in autophagy; however, the detailed mechanism of its activity remains unclear. Here, we used a Systematic Evolution of Ligands by EXponential enrichment (SELEX) screening approach to identify an RNA aptamer of 40 nucleotides that specifically recognizes and binds to intracellular lysosomal Ptdlns(3)P. Binding occurs in a magnesium concentration- and pH-dependent manner, and consequently inhibits autophagy as determined by LC3II/I conversion, p62 degradation, formation of LC3 puncta, and lysosomal accumulation of Phafin2. These effects in turn inhibited lysosomal acidification, and the subsequent hydrolytic activity of cathepsin D following induction of autophagy. Given the essential role of Ptdlns(3)P as a key targeting molecule for autophagy induction, identification of this novel Ptdlns(3)P RNA aptamer provides new opportunities for investigating the biological functions and mechanisms of phosphoinositides. (C) 2019 The Authors. Published by Elsevier Inc.