ABSTRACT

N⁶-methyladenosine (m⁶A) has emerged as a crucial epitranscriptomic mark which regulates a broad spectrum of physiological processes including stem cell differentiation. m⁶A-binding YTHDF proteins have recently been proposed to mediate differentiation of leukemia cell in a redundant manner. However, whether these proteins play semblable roles in pluripotent stem cell remain largely unknown. Here, we showed the differential functions of YTHDF1 and YTHDF3 in controlling the differentiation of embryonic stem cells (ESCs). Depletion of YTHDF3 in ESCs resulted in loss of pluripotency with accelerated expressions of marker genes involved in formation of three germ layers. Phenotypic and transcriptomic analyses revealed that loss of YTHDF1 led to dramatic impairment of cardiomyocytes (CMs) differentiation, accompanied by downregulated CM-specific genes. While, knockdown of YTHDF3 accelerated differentiation through facilitating the expressions of CM-specific gene. Notably, YTHDF3 appears to modulate cellular differentiation partially through suppression of YTHDF1, supporting the distinguishable but interrelated roles of YTHDF1 and YTHDF3 in cell fate determination.

摘要

N ^6-甲基腺苷（m ⁶ A）已成为一种重要的转录组标记，可调节包括干细胞分化在内的各种生理过程。⁶米最近已经提出了A结合型YTHDF蛋白以冗余方式介导白血病细胞的分化。然而，这些蛋白在多能干细胞中是否起着相似的作用仍是未知之数。在这里，我们显示了YTHDF1和YTHDF3在控制胚胎干细胞（ESC）分化中的差异功能。胚胎干细胞中YTHDF3的耗尽导致多能性的丧失，并加速了参与形成三个胚层的标记基因的表达。表型和转录组学分析表明，YTHDF1的丧失导致心肌细胞（CMs）分化急剧受损，并伴有CM特异性基因下调。同时，YTHDF3的敲低通过促进CM特异性基因的表达来加速分化。值得注意的是