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哺乳动物RNA外切核酸酶5/NEF-sp作为睾丸特异性核3'→5'外切核糖核酸酶的特性分析。

Characterization of the mammalian RNA exonuclease 5/NEF-sp as a testis-specific nuclear 3' → 5' exoribonuclease.

作者信息

Silva Sara, Homolka David, Pillai Ramesh S

机构信息

Department of Molecular Biology, University of Geneva, CH-1211 Geneva 4, Switzerland.

European Molecular Biology Laboratory, Grenoble Outstation, 38042, France.

出版信息

RNA. 2017 Sep;23(9):1385-1392. doi: 10.1261/rna.060723.117. Epub 2017 May 24.

DOI:10.1261/rna.060723.117
PMID:28539487
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5558908/
Abstract

Ribonucleases catalyze maturation of functional RNAs or mediate degradation of cellular transcripts, activities that are critical for gene expression control. Here we identify a previously uncharacterized mammalian nuclease family member NEF-sp (RNA exonuclease 5 [REXO5] or LOC81691) as a testis-specific factor. Recombinant human NEF-sp demonstrates a divalent metal ion-dependent 3' → 5' exoribonuclease activity. This activity is specific to single-stranded RNA substrates and is independent of their length. The presence of a 2'--methyl modification at the 3' end of the RNA substrate is inhibitory. Ectopically expressed NEF-sp localizes to the nucleolar/nuclear compartment in mammalian cell cultures and this is dependent on an amino-terminal nuclear localization signal. Finally, mice lacking NEF-sp are viable and display normal fertility, likely indicating overlapping functions with other nucleases. Taken together, our study provides the first biochemical and genetic exploration of the role of the NEF-sp exoribonuclease in the mammalian genome.

摘要

核糖核酸酶催化功能性RNA的成熟或介导细胞转录本的降解，这些活性对于基因表达控制至关重要。在这里，我们鉴定出一种以前未被表征的哺乳动物核酸酶家族成员NEF-sp（RNA外切核酸酶5 [REXO5]或LOC81691）作为睾丸特异性因子。重组人NEF-sp表现出二价金属离子依赖性的3'→5'核糖核酸外切酶活性。这种活性对单链RNA底物具有特异性，并且与其长度无关。RNA底物3'端存在2'-甲基修饰具有抑制作用。在哺乳动物细胞培养物中异位表达的NEF-sp定位于核仁/核区室，这取决于氨基末端核定位信号。最后，缺乏NEF-sp的小鼠是可存活的，并且表现出正常的生育能力，这可能表明其与其他核酸酶功能重叠。综上所述，我们的研究首次对NEF-sp核糖核酸外切酶在哺乳动物基因组中的作用进行了生化和遗传学探索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/5d56c92ef0be/1385f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/e88901f418e8/1385f01.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/a4e1da327560/1385f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/5d56c92ef0be/1385f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/e88901f418e8/1385f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/339b6e666f43/1385f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/a4e1da327560/1385f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b82/5558908/5d56c92ef0be/1385f04.jpg

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