ADBP-1 Regulates an ADAR RNA-Editing Enzyme to Antagonize RNA-Interference-Mediated Gene Silencing in Caenorhabditis elegans
ADBP-1 Regulates an ADAR RNA-Editing Enzyme to Antagonize RNA-Interference-Mediated Gene Silencing in Caenorhabditis elegans
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DOI：
10.1534/genetics.108.093310
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发表时间：
2008-10-01
期刊：
影响因子：
3.3
通讯作者：
Ishihara, Takeshi
中科院分区：
文献类型：
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作者：
Ohta, Hiromitsu;Fujiwara, Manabi;Ishihara, Takeshi
小干扰RNA（siRNA）和微小RNA（miRNA）通过进化上保守的途径介导基因沉默。在秀丽隐杆线虫中，也已知siRNA/miRNA途径影响转基因表达。为了鉴定调节siRNA/miRNA途径效率的基因，我们使用转基因的表达水平作为基因沉默的指标，并分离出转基因沉默突变体adbp-1（ADR-2结合蛋白）。adbp-1突变导致转基因沉默在皮下和肠细胞中的细胞自主的方式，这取决于RNA干扰（RNAi）机制。adbp-1基因编码的蛋白质没有保守的结构域，定位在细胞核中。酵母双杂交筛选和免疫共沉淀分析表明，ADBP-1与ADR-2发生物理相互作用，ADR-2是来自阿达尔（腺苷脱氨酶作用于dsRNA）家族的RNA编辑酶。在adbp-1突变体中，如先前在adr-2突变体中所示，未检测到A至I RNA编辑，表明ADBP-1是ADR-2的RNA编辑活性所必需的。我们发现ADBP-1促进ADR-2的核定位。ADBP-1可能调节ADR-2活性和随后的RNA编辑，从而拮抗RNAi介导的转基因沉默。优雅
Small interfering RNAs (siRNAs) and microRNAs (miRNAs) mediate gene silencing through evolutionarily conserved pathways. In Caenorhabditis elegans, the siRNA/miRNA pathways are also known to affect transgene expression. To identify genes that regulate the efficiencies of the siRNA/miRNA pathways, we used the expression level of a transgene as an indicator of gene silencing and isolated a transgene-silencing mutant, adbp-1 (ADR-2 binding protein). The adbp-1 mutation caused transgene silencing in hypodermal and intestinal cells in a cell-autonomous manner, depending on the RNA interference (RNAi) machinery. The adbp-1 gene encodes a protein with no conserved domains that is localized in the nucleus. Yeast two-hybrid screening and co-immunoprecipitation analysis demonstrated that ADBP-1 physically interacts with ADR-2, an RNA-editing enzyme from the ADAR (adenosine deaminase acting on dsRNA) family. In the adbp-1 mutant, as previously shown in adr-2 Mutants, A-to-I RNA editing was not detected, suggesting that ADBP-1 is required for the RNA-editing activity of ADR-2. We found that ADBP-1 facilitates the nuclear localization of ADR-2. ADBP-1 may regulate ADR-2 activity and the consequent RNA editing and thereby antagonize RNAi-mediated transgene silencing in C. elegans.