Dataset Information

LIN28a modulates ESC differentiation by interacting with PRC2

ABSTRACT: LIN28a is an RNA binding protein that was initially characterized for its role in the negative regulation of let-7 microRNA biogenesis. In the last years a new role in the regulation of mRNA translation has been highlighted. This mechanism seems to be particularly relevant in the exit of embryonic stem cells (ESCs) from the naïve state and in the first phases of differentiation process. We have demonstrated that in the first step of ESC differentiation, i.e. the establishment of epiblast-like-cells and the formation of neural precursors, LIN28a enhances the translation of de novo DNA methyltransferase Dnmt3a [1] and blocks that of the chromatin architectural factor Hmga2 [2]. How LIN28a fulfils these opposite functions in ESC differentiation is still unknown. Our preliminary results indicated that LIN28a contributes in several ways to guide ESC differentiation possibly by changing molecular partners and regulating different targets. To explore this idea, we have generated Lin28a KO ESCs using CRISPR-Cas9 technology to study how Lin28a contributes to ESC differentiation. We have found that Lin28a KO ESCs are not able to properly exit from the naïve state to undergo a differentiation program. This effect seems to be due to the impairment of different cellular processes. To better characterize this role in ESC fate we have analyzed the molecular interactions of LIN28a at different steps of ESC differentiation through proteomics approaches. Moreover, to identify the direct mRNA targets of LIN28a we are developing an iCLIP protocol suitable to analyze the binding of LIN28a to RNAs during ESC differentiation. These data coupled with the phenotypic and proteomics analysis will contribute to understand the emerging fundamental role of RNA binding proteins in differentiation and development.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryonic Stem Cell

SUBMITTER: CHIARA D'AMBROSIO

LAB HEAD: Andrea Scaloni

PROVIDER: PXD049332 | Pride | 2026-05-05

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 185

Similar Datasets

Project description:At the blastocyst stage, the embryo invades the uterine and the pluripotent epiblast cells get reshaped from an amorphous ball into a polarized cup-shaped epithelium. At the same time, the transiently established naïve pluripotency is dismantled, transforming into a more developmentally advanced post-implantation pluripotent state. The major driver of the differentiation process is the autocrine Fgf/Mek/Erk singling which promotes the exit of naïve pluripotency. However, inhibiting Mek alone is not sufficient to maintain the cells in the naïve state suggesting the contribution of additional, as yet unidentified, pro-differentiation signals. Using a 3D embryonic stem cells (ESC) based model of epiblast development we established an automated pipeline for cell culture and analysis and performed a large-scale functional screen to identify factors regulating the development of the pluripotent lineage. We found that the mitotic kinases – Aurora kinase A (Aurka) and Polo-like kinase (PLK) positively regulate Erk signalling in a Mek-independent manner. We determined that Aurka and Erk form a complex in ESCs and suppression of Aurka activity delays the exit of naïve pluripotency. Moreover, a combination of Aurka and Mek inhibition enabled the de novo derivation of ESC lines from mouse embryos. Interestingly, during cell division, we found that phospho-Erk (pErk) localizes on the spindle poles at prophase and metaphase, gradually decreasing its signal at anaphase and telophase. Proximity biotinylation analysis of the pErk interactome identified the formation of PLK/pErk complex in metaphase cells. Moreover, inhibition of PLK, but not Mek or Aurka, resulted in a substantial decrease in Erk phosphorylation at the mitotic spindle. In summary, our findings demonstrate that the pluripotent cells continuously generate and perceive two types of Erk-mediated pro-differentiation cues - autocrine signals of the Fgf/Mek cascade and cell-intrinsic cues via the mitotic kinase Aurka and PLK. Altogether, these signals promote the exit of naïve pluripotency, driving the transient pace of embryonic development.

			Action	DRS
	IND_LIN28_.msf	Msf
	IND_LIN28_1.raw	Raw
	IND_LIN28_10.raw	Raw
	IND_LIN28_11.raw	Raw
	IND_LIN28_12.raw	Raw

Dataset Information

LIN28a modulates ESC differentiation by interacting with PRC2

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets