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LINE-1 ORF1 protein enhances Alu SINE retrotransposition.

机译:LINE-1 ORF1蛋白增强Alu SINE逆转座。

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摘要

Retroelements have contributed over one third of the human genome mass. The currently active LINE-1 (L1) codes for two proteins (ORF1p and ORF2p), both strictly required for retrotransposition. In contrast, the non-coding parasitic SINE (Alu) only appears to need the L1 ORF2p for its own amplification. This requirement was previously determined using a tissue culture assay system in human cells (HeLa). Because HeLa are likely to express functional L1 proteins, it is possible that low levels of endogenous ORF1p are necessary for the observed tagged Alu mobilization. By individually expressing ORF1 and ORF2 proteins from both human (L1RP and LRE3) and rodent (L1A102 and L1spa) L1 sources, we demonstrate that increasing amounts of ORF1 expressing vector enhances tagged Alu mobilization in HeLa cells. In addition, using chicken fibroblast cells as an alternate cell culture source, we confirmed that ORF1p is not strictly required for Alu mobilization in our assay. Supporting our observations in HeLa cells, we find that tagged Alu retrotransposition is improved by supplementation of ORF1p in the cultured chicken cells. We postulate that L1 ORF1p plays either a direct or indirect role in enhancing the interaction between the Alu RNA and the required factors needed for its retrotransposition.
机译:反义元素贡献了人类基因组质量的三分之一。当前活跃的LINE-1(L1)编码两种蛋白质(ORF1p和ORF2p),这两种蛋白质都是逆转座所必需的。相反,非编码寄生SINE(Alu)似乎只需要L1 ORF2p即可进行自身的放大。先前使用人体细胞中的组织培养测定系统(HeLa)确定了这一要求。由于HeLa可能表达功能性L1蛋白,因此对于观察到的标记Alu动员,可能需要低水平的内源性ORF1p。通过分别从人(L1RP和LRE3)和啮齿动物(L1A102和L1spa)L1来源表达ORF1和ORF2蛋白,我们证明增加表达量的ORF1载体可增强HeLa细胞中标记的Alu动员。此外,使用鸡成纤维细胞作为备用细胞培养源,我们证实在我们的测定中,ORF1p并不是Alu动员的严格要求。支持我们在HeLa细胞中的观察,我们发现通过在培养的鸡细胞中补充ORF1p可以改善标记的Alu逆转座。我们假设L1 ORF1p在增强Alu RNA及其逆转所需的必需因子之间的相互作用中起直接或间接的作用。

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