この研究では、幹細胞が自己複製を行うために必要な分子イベントの複雑なメカニズムに焦点を当て、SOX2転写因子の動態を明らかにしています。特に、単一分子トラッキング技術とSTREAMING-tag転写レポーターシステムを用いて、胚性幹細胞におけるNanog遺伝子座でのSOX2のクラスター形成パターンと、転写バーストとの関係をリアルタイムで可視化しました。この研究により、SOX2のクラスター形成が転写バーストと異なるタイミングで起こることが示され、SOX2が転写サイクルの各段階で異なる役割を果たしていることがわかりました。
https://www.biorxiv.org/content/10.1101/2024.09.10.612363v1
Anti-phase clustering of regulatory factors shapes gene bursting
Bitong Li, Yew Yan Wong, Neftali Flores-Rodriguez, Tara Davidson, Matthew S Graus, Valeriia Smialkovska, Hiroaki Ohishi, Angelika Feldmann, Hiroshi Ochiai, Mathias Francois
Abstract
The ability of stem cells to divide and self-renew depends on a complex choreography of molecular events that maintain the transcriptional oscillation of pluripotency genes. Only a handful of transcription factors (TFs) are necessary to preserve pluripotency and reprogram differentiated cells into stem cells. Paradoxically, while the protein players are known, the challenge remains to decipher the series of steps that TFs undertake to modulate on and off fluctuations of gene transcription. Here, we use single-molecule tracking combined with the STREAMING-tag transcriptional reporter systems to reveal temporal clustering patterns of endogenous SOX2 occupancy at the Nanog locus in relation to its nascent mRNA synthesis in live embryonic stem cells. These patterns distinctively outline multifaceted regulatory behaviours of SOX2 associated with various stages of the Nanog transcription cycle. This study exposes that SOX2 clustering activity is out-of-phase with regulatory factors that engage with transcription burst at the Nanog gene locus.