Department of Biochemistry, Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany and Friedrich Miescher Laboratory of the Max Planck Society, Spemannstrasse 39, 72076 Tübingen, Germany.
Nucleic Acids Res. 2013 Dec;41(22):10563-72. doi: 10.1093/nar/gkt786. Epub 2013 Sep 3.
Non-LTR retrotransposons are mobile genetic elements and play a major role in eukaryotic genome evolution and disease. Similar to retroviruses they encode a reverse transcriptase, but their genomic integration mechanism is fundamentally different, and they lack homologs of the retroviral nucleocapsid-forming protein Gag. Instead, their first open reading frames encode distinct multi-domain proteins (ORF1ps) presumed to package the retrotransposon-encoded RNA into ribonucleoprotein particles (RNPs). The mechanistic roles of ORF1ps are poorly understood, particularly of ORF1ps that appear to harbor an enzymatic function in the form of an SGNH-type lipolytic acetylesterase. We determined the crystal structures of the coiled coil and esterase domains of the ORF1p from the Danio rerio ZfL2-1 element. We demonstrate a dimerization of the coiled coil and a hydrolytic activity of the esterase. Furthermore, the esterase binds negatively charged phospholipids and liposomes, but not oligo-(A) RNA. Unexpectedly, the esterase can split into two dynamic half-domains, suited to engulf long fatty acid substrates extending from the active site. These properties indicate a role for lipids and membranes in non-LTR retrotransposition. We speculate that Gag-like membrane targeting properties of ORF1ps could play a role in RNP assembly and in membrane-dependent transport or localization processes.
非长末端重复转座子是移动遗传元件,在真核生物基因组进化和疾病中发挥着重要作用。它们与逆转录病毒相似,编码逆转录酶,但它们的基因组整合机制截然不同,并且缺乏逆转录病毒核衣壳形成蛋白 Gag 的同源物。相反,它们的第一个开放阅读框编码独特的多结构域蛋白(ORF1ps),推测将逆转座子编码的 RNA 包装到核糖核蛋白颗粒(RNP)中。ORF1ps 的作用机制了解甚少,特别是那些似乎具有 SGNH 型脂肪酶乙酰酯酶形式的酶促功能的 ORF1ps。我们确定了来自斑马鱼 ZfL2-1 元件的 ORF1p 的卷曲螺旋和酯酶结构域的晶体结构。我们证明了卷曲螺旋的二聚化和酯酶的水解活性。此外,酯酶结合带负电荷的磷脂和脂质体,但不结合寡聚(A)RNA。出乎意料的是,酯酶可以分裂成两个动态的半结构域,适合吞噬从活性位点延伸的长脂肪酸底物。这些特性表明脂质和膜在非长末端重复转座中起作用。我们推测 ORF1ps 类似 Gag 的膜靶向特性可能在 RNP 组装以及膜依赖的运输或定位过程中发挥作用。