Abstract
Peg10 (paternally expressed gene 10) is an imprinted gene that is essential for placental development. It is thought to derive from a Ty3-gyspy LTR (long terminal repeat) retrotransposon and retains Gag and Pol-like domains. Here we show that the Gag domain of PEG10 can promote vesicle budding similar to the HIV p24 Gag protein. Expressed in a subset of mouse endocrine organs in addition to the placenta, PEG10 was identified as a substrate of the deubiquitinating enzyme USP9X. Consistent with PEG10 having a critical role in placental development, PEG10-deficient trophoblast stem cells (TSCs) exhibited impaired differentiation into placental lineages. PEG10 expressed in wild-type, differentiating TSCs was bound to many cellular RNAs including Hbegf (Heparin-binding EGF-like growth factor), which is known to play an important role in placentation. Expression of Hbegf was reduced in PEG10-deficient TSCs suggesting that PEG10 might bind to and stabilize RNAs that are critical for normal placental development.
Highlights
Transposable elements (TEs) are one of the biggest threats to the integrity of prokaryotic and eukaryotic genomes because their insertion into coding or regulatory regions could disrupt essential genes [1,2,3]
PEG10 is regulated by USP9X
We identified PEG10 while looking for substrates of the deubiquitinating enzyme USP9X in mouse embryonic stem cells (ESCs)
Summary
Transposable elements (TEs) are one of the biggest threats to the integrity of prokaryotic and eukaryotic genomes because their insertion into coding or regulatory regions could disrupt essential genes [1,2,3]. We study the role of PEG10 in mouse embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) after identifying it as a substrate of the deubiquitinating enzyme USP9X. Cells were fixed with 8% PFA, blocked for 1 h with 10% donkey serum, 2% BSA, and 0.2% saponin in PBS, incubated overnight at 4 ̊C in 5 μg/ml anti-PEG10 antibody in blocking buffer, and for 1 h at room temperature in 2 μg/mL Alexa 488-conjugated donkey antirat antibody (A-21208, Thermo Fisher).
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