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反转录转座子衍生的人类 PEG10 蛋白酶的功能研究。

Functional Study of the Retrotransposon-Derived Human PEG10 Protease.

机构信息

Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary.

Doctoral School of Molecular Cell and Immune Biology, University of Debrecen, 4032 Debrecen, Hungary.

出版信息

Int J Mol Sci. 2020 Mar 31;21(7):2424. doi: 10.3390/ijms21072424.

DOI:10.3390/ijms21072424
PMID:32244497
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7212762/
Abstract

Paternally expressed gene 10 (PEG10) is a human retrotransposon-derived imprinted gene. The mRNA of encodes two protein isoforms: the Gag-like protein (RF1) is coded by reading frame 1, while the Gag-Pol-like polyprotein (RF1/RF2) is coded by reading frames 1 and 2. The proteins are translated by a typical retroviral frameshift mechanism. The protease (PR) domain of RF2 contains an -Asp-Ser-Gly- sequence, which corresponds to the consensus -Asp-Ser/Thr-Gly- active-site motif of retroviral aspartic proteases. The function of the aspartic protease domain of RF2 remains unclear. To elucidate the function of PEG10 protease (PR), we designed a frameshift mutant (RF1/RF2) for comparison with the RF1/RF2 form. To study the effects of PR on cellular proliferation and viability, mammalian HEK293T and HaCaT cells were transfected with plasmids coding for either RF1/RF2, the frameshift mutant (RF1/RF2), or a PR active-site (D370A) mutant RF1/RF2. Our results indicate that RF1/RF2 overexpression results in increased cellular proliferation. Remarkably, transfection with RF1/RF2 had a detrimental effect on cell viability. We hypothesize that PR plays an important role in the function of this retroviral remnant, mediating the proliferation of cells and possibly implicating it in the inhibition of apoptosis.

摘要

父系表达基因 10（PEG10）是一种人类反转录转座子衍生的印迹基因。该基因的 mRNA 编码两种蛋白异构体：Gag 样蛋白（RF1）由阅读框 1 编码，而 Gag-Pol 样多蛋白（RF1/RF2）由阅读框 1 和 2 编码。这些蛋白通过典型的逆转录病毒移码机制翻译。RF2 的蛋白酶（PR）结构域含有一个 -Asp-Ser-Gly- 序列，该序列与逆转录病毒天冬氨酸蛋白酶的保守 -Asp-Ser/Thr-Gly- 活性位点模体相对应。RF2 的天冬氨酸蛋白酶结构域的功能尚不清楚。为了阐明 PEG10 蛋白酶（PR）的功能，我们设计了一个移码突变体（RF1/RF2）与 RF1/RF2 形式进行比较。为了研究 PR 对细胞增殖和活力的影响，用编码 RF1/RF2、移码突变体（RF1/RF2）或 PR 活性位点（D370A）突变体 RF1/RF2 的质粒转染哺乳动物 HEK293T 和 HaCaT 细胞。我们的结果表明，RF1/RF2 的过表达导致细胞增殖增加。值得注意的是，RF1/RF2 的转染对细胞活力有不利影响。我们假设 PR 在这个逆转录病毒残余物的功能中发挥重要作用，介导细胞的增殖，并可能使其参与抑制细胞凋亡。

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