SAF-A和nuclear RNA形成网络调控转录

Nuclear RNA forms an interconnected network of transcription-dependent and tunable microgels (biorxiv.org)

科学问题:细胞核内的微环境是怎样的

提出假设:SAF-A和nuclear RNA形成一种胶状网络结构,文中成为”microgel”。microgel内部蛋白质的扩散速率更低,而microgel多的地方的染色质更松散、转录更活跃。microgel本身可以被一种核酸内切酶XRN2降解。

主要实验:

1)通过超分辨显微镜观察到SAF-A和nuclear RNA形成microgel结构

2)确定了SAF-A中跟RNA相互作用的结构域

3)KD SAF-A或抑制转录后microgel减少了

4)伴随着上图中microgel的减少,核内蛋白质的扩散效率增强了

5)microgel本身可以被一种核酸内切酶XRN2降解

图G对交联后对SAF-A做WB,当α-amanitin抑制转录,同时存在XRN2时,SAF-A更多是monomer.

最后还有一些figure研究了micro对染色质“去压缩”的功能。

总结:本文研究揭示了RNA和SAF-A相互作用形成的微凝胶在细胞核内的重要作用。通过高分辨率成像和分子动力学模拟,作者发现这些微凝胶促进染色质去压缩,从而调控基因表达。这一机制通过RNA降解酶XRN2的调控进一步得到了验证。

一些Q&A.

1)目光怎么转移到XRN2上的?

答:通过BioID技术发现XRN2是SAF-A的proximal proteins. 结合XRN2是一种核酸内切酶,于是联想到它是否能通过降解nuclear RNA来调控microgel.

We reasoned that proteins important for RNA turnover were likely to be spatially located in the vicinity of the nucleoplasmic network and microgels, so BioID74 (proximity-dependent biotin identification) was used to identify proteins in the vicinity of the SAF-A, by tagging the protein on the N-terminus with BirA. As expected, SAF-A was observed to be in close proximity to its homologues HNRNPUL1 and HNRNPUL2 and other members of a nuclear scaffold: SAFB, SAFB2 and MATRIN3 (Fig S4A; Table S1), but, surprisingly, 5′-3′ Exoribonuclease 2 (XRN2) was also enriched.