RYBP regulates Pax6 during in vitro neural differentiation of mouse embryonic stem cells

Enikő Sutus,Gergő Kovács,Surya Henry,Lili Adorján,Melinda Katalin Pirity

doi:10.1038/s41598-022-06228-1

Enikő Sutus, Gergő Kovács + Show 3 more

Open Access

https://doi.org/10.1038/s41598-022-06228-1

Copy DOI

Abstract

We have previously reported that RING1 and YY1 binding protein (RYBP) is important for central nervous system development in mice and that Rybp null mutant (Rybp−/−) mouse embryonic stem (ES) cells form more progenitors and less terminally differentiated neural cells than the wild type cells in vitro. Accelerated progenitor formation coincided with a high level of Pax6 expression in the Rybp−/− neural cultures. Since Pax6 is a retinoic acid (RA) inducible gene, we have analyzed whether altered RA signaling contributes to the accelerated progenitor formation and impaired differentiation ability of the Rybp−/− cells. Results suggested that elevated Pax6 expression was driven by the increased activity of the RA signaling pathway in the Rybp−/− neural cultures. RYBP was able to repress Pax6 through its P1 promoter. The repression was further attenuated when RING1, a core member of ncPRC1s was also present. According to this, RYBP and PAX6 were rarely localized in the same wild type cells during in vitro neural differentiation. These results suggest polycomb dependent regulation of Pax6 by RYBP during in vitro neural differentiation. Our results thus provide novel insights on the dynamic regulation of Pax6 and RA signaling by RYBP during mouse neural development.

Highlights

We have previously reported that RING1 and Yin-yang-1 transcription factor (YY1) binding protein (RYBP) is important for central nervous system development in mice and that Rybp null mutant (Rybp−/−) mouse embryonic stem (ES) cells form more progenitors and less terminally differentiated neural cells than the wild type cells in vitro
There were 7 genes upregulated in Rybp−/− ES cells including Alcohol dehydrogenase 4 (Adh4), Retinol binding protein 7 (Rbp7), Alcohol dehydrogenase 7 (Adh7), Retinoic acid receptor beta (Rarβ), Cellular retinoic acid binding protein (Crabp1), Retinol binding protein 4 (Rbp4), Aldehyde dehydrogenase 1 family member B1 (Aldh1b1) and 3 genes were downregulated including Pax[6], Cytochrome P450 Family 26 Subfamily A Member 1 (Cyp26a1) and Signaling receptor and transporter of retinol (Stra6)
Since Pax[6] is robustly activated by RA signaling during neural development, we examined whether the expression of RA signaling pathway members is attenuated after alltrans retinoic acid treatment and whether the Rybp−/− neural cultures express more Pax[6] than the wild type cells

Summary

Introduction

We have previously reported that RING1 and YY1 binding protein (RYBP) is important for central nervous system development in mice and that Rybp null mutant (Rybp−/−) mouse embryonic stem (ES) cells form more progenitors and less terminally differentiated neural cells than the wild type cells in vitro. Since Pax[6] is a retinoic acid (RA) inducible gene, we have analyzed whether altered RA signaling contributes to the accelerated progenitor formation and impaired differentiation ability of the Rybp−/− cells. RYBP and PAX6 were rarely localized in the same wild type cells during in vitro neural differentiation. In case of mouse embryonic stem (ES) cells RA induction facilitates neural differentiation in a concentration dependent manner and the differentiation of ES cells correlates with increased activity of Paired box 6 (Pax6), transcription factor, a gene important in central nervous system d evelopment[4]. Both PRC1s and ncPRC1s contain Ring finger proteins (RING1, RNF2) and Polycomb group ring finger catalytic (PCGF) core, but in ncPRC1s, RING1

Objectives

Methods

Results

Conclusion