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Pcbp1 constrains Oct4 expression in the context of pluripotency

Bakhmet, E. I.; Zinovyeva, A. S.; Kuzmin, A. A.; Smirnova, D. V.; Gordeev, M. N.; Petrenko, E. E.; Aksenov, N. D.; Tomilin, A. N.

2024-09-08 cell biology
10.1101/2024.09.07.611681 bioRxiv
Show abstract

Oct4 is a commonly known marker of pluripotent stem cells as well as one of the key factors required for pluripotency induction. Its gene (Pou5f1) is subject to complicated regulation through distal and proximal enhancers. Noteworthy, this protein also plays an important role in primitive endoderm (PrE) specification, though the mechanisms driving its expression during this process are still unknown. Here we show that KH-domain protein Pcbp1 occupies poly(C)- sites of the Pou5f1 enhancers, but Pcbp1 knockout does not affect the Oct4 expression level in ESCs. On the contrary, Pcbp1 is essential for timely Oct4 downregulation upon differentiation signals. Indeed, Pcbp1 loss results in high rate of spontaneous differentiation of ESCs to PrE, and this effect is enhanced upon retinoic acid treatment. This phenotype can be explained by residual Oct4 expression, as Oct4 depletion rescues normal differentiation. Overall, our results point to Pcbp1 is a transcriptional regulator of Pou5f1, purported to synchronize Oct4 expression decline with the pluripotency network shutdown during differentiation. Oct4 being outside of this network loss its functions as factor of pluripotency and acts as PrE specifier.

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