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Nature Cell Biology:江浩团队揭示AKAP95可通过相分离调控基因剪接和肿瘤生成

2020-07-28 BioArt

癌症常与基因表达紊乱有关, 包括在染色体、转录、mRNA剪接等水平上的调控异常。就像所有细胞内的化学反应,基因表达并非均匀分布在细胞核内,而是在时空上被分隔化了才得以正确进行和调控。

癌症常与基因表达紊乱有关, 包括在染色体、转录、mRNA剪接等水平上的调控异常。就像所有细胞内的化学反应,基因表达并非均匀分布在细胞核内,而是在时空上被分隔化了(compartmentalized)才得以正确进行和调控。除了有膜结构的细胞器,另一类无膜结构的细胞器也是很重要的分隔化手段。近几年的研究显示液液相分离(liquid-liquid phase separation) 是形成所谓的生物分子凝聚体(biomolecular condensates) 和许多无核细胞器的根本推动力。配合不同的生物过程,这些凝聚体有着一系列不同的生物物理属性,包括从分子高度能动的液体到半流体或胶质到分子能动性很差的纤维和固体状态,包括染色体、转录、以及mRNA剪接的分子都能形成凝聚体而有活性。然而,目前并不知道凝聚体的不同生物物理属性对基因表达以及癌症有何影响。

2020年7月27日,弗吉尼亚大学江浩团队在Nature Cell Biology杂志上发表文章Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis,揭示了核内蛋白AKAP95在合适的凝聚体生物物理属性的范围内可通过液液相分离调控基因剪接和肿瘤生成。

研究人员在前期研究发现核内蛋白AKAP95能够调控基因表达, 特别是mRNA剪接。在这项研究中,他们发现AKAP95直接调控一些癌症基因的mRNA剪接从而支持癌症细胞的生长。小鼠敲除Akap95基因后,其发育和生理都没影响。这些小鼠的胚胎成纤维细胞生长正常,但很难被转化成癌细胞, 而且在癌基因活跃时进入细胞衰老(senescence), 伴随着mRNA剪切异常。所以AKAP95可能是潜在的癌症治疗靶点。

接下来他们研究这个蛋白调控剪接的分子机制。由于常观察到该蛋白在变性条件下异乎寻常的自结合能力, 发现了AKAP95通过相分离在体外形成液状微粒和在核内的高度能动的凝聚体。将它一段内在无序区(Intrinsically Disordered Region, IDR)的酪氨酸 (Tyr) 突变成丙氨酸(Ala)或丝氨酸(Ser)使它完全失去分相能力, 并且使它失去了剪接调控的能力。将它自身的IDR替换成来自一些不相关蛋白的具有相分离能力的IDR则能使它保持分相以及剪接调控的能力。这些结果支持AKAP95需要相分离能力来调控剪接。

有意思的是IDR的酪氨酸突变成苯丙氨酸(Phe)的一个对照(因为这两个氨基酸都有芳香环)。这个突变体也形成凝聚体, 但是其剪接调控的能力却显着下降。这引起了作者的注意。通过仔细的定量研究, 他们发现这个突变体分相能力更强,体外和胞内形成的凝聚体更倾向于固态而非液态,凝聚体内的分子能动性和扩散系数显着下降(和UCI Gratton 团队的合作)。这样这个突变就减弱了剪接反应中的分子运动和碰撞, 从而减弱了剪接反应的效率。

最后他们回到了人和小鼠细胞系统去检查AKAP95相分离对癌细胞以及内源基因剪接的调控。他们发现, 失去相分离能力的突变体完全失去了支持癌症细胞生长并抑制它们senescence的能力以及内源基因剪接的调控能力, 而使凝聚体固化的突变也显着减弱了所有这些能力。

总之,这项研究揭示了一些基因调控蛋白如AKAP95等不仅需要相分离形成凝聚体, 还需要在一个合适的凝聚体生物物理属性的范围内才有好的活性来调控基因表达以及癌症。这也为癌症治疗提供了一个非常规的思路---也许可以通过对凝聚体属性的两个方向进行干预(化掉或硬化)来抑制癌细胞。

原始出处:

Wei Li, Jing Hu, Bi Shi, et al.Biophysical properties of AKAP95 protein condensates regulate splicing and tumorigenesis.Nature Cell Biology (2020).Published: 27 July 2020

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    2021-06-08 anan
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    2021-04-28 sunylz
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    2020-09-13 liye789132251
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    2020-10-25 维他命
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