Functional Study of the Retrotransposon-Derived Human PEG10 Protease

Abstract

Paternally expressed gene 10 (PEG10) is a human retrotransposon-derived imprinted gene. The mRNA of PEG10 encodes two protein isoforms: the Gag-like protein (RF1_PEG10) is coded by reading frame 1, while the Gag-Pol-like polyprotein (RF1/RF2_PEG10) is coded by reading frames 1 and 2. The proteins are translated by a typical retroviral frameshift mechanism. The protease (PR) domain of RF2_PEG10 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_PEG10 remains unclear. To elucidate the function of PEG10 protease (PR_PEG10), we designed a frameshift mutant (_fsRF1/RF2_PEG10) for comparison with the RF1/RF2_PEG10 form. To study the effects of PR_PEG10 on cellular proliferation and viability, mammalian HEK293T and HaCaT cells were transfected with plasmids coding for either RF1/RF2_PEG10, the frameshift mutant (_fsRF1/RF2_PEG10), or a PR active-site (D370A) mutant _fsRF1/RF2_PEG10. Our results indicate that _fsRF1/RF2_PEG10 overexpression results in increased cellular proliferation. Remarkably, transfection with _fsRF1/RF2_PEG10 had a detrimental effect on cell viability. We hypothesize that PR_PEG10 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.

Keywords: PEG10; cell proliferation; cell viability; cis protease activity; homology modeling; paternally expressed gene 10; protease; retrotransposon; retroviral-like protease; ubiquitination.