酸-3-酸盐通过多环体路径调解弧囊分泌
Kritika Mehta1,2, Henry Yentsch1, Jungbin Lee1
1Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL 61801.
概括
酸-3-酸盐 (PI3P) 通过内体-多细胞体路径调节活性细胞骨相关蛋白 (Arc) 囊体的组合和分泌,影响RNA转移和神经元可塑性.
科学领域:
- 神经科学是一个神经科学.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 活动调节的细胞骨相关蛋白 (Arc/Arg3.1) 对学习和记忆至关重要.
- 弧形蛋白通过类似病毒的囊促进细胞间RNA转移,但其分泌途径尚不清楚.
- 弧形与病毒群特异抗原 (Gag) 蛋白具有结构上的相似性.
研究的目的:
- 为了阐明弧形虫的调节器和分泌途径.
- 为了调查酸-3-酸盐 (PI3P) 在Arc酸组装和贩运中的作用.
- 了解Arc介导细胞间RNA转移的机制.
主要方法:
- 用PI3P进行Arc蛋白和体外结合试验的复制.
- 在HEK293T细胞中进行共聚焦和超分辨率显微镜,以可视化使用内体标记器 (FYVE,Rab5,CD63) 的Arc局部化.
- 编辑CRISPR/Cas9基因以淘汰RalA和RalBGTPases;评估RNA转移效率和神经元树突脊柱形态;评估小鼠皮层神经元中的细胞外囊泡吸收.
主要成果:
- 复制的 Arc 蛋白质优先结合到 PI3P.
- 弧点点与PI3P,早期内体和多体细胞体 (MVB) 标记物结合,在MVB内囊中积累.
- 拉拉/B的CRISPR淘汰显著减少了Arc介导的RNA转移,并在培养的神经元中增加了成熟的树突.
- 从Arc表达细胞中吸收细胞外囊泡降低了小鼠皮层神经元中的表面GlutA1水平,这种效应在RalA/B敲击中被取消.
结论:
- 酸-3-酸盐 (PI3P) 通过内体-MVB通路调解状体的组合和分泌.
- 弧形囊组合是由内细胞膜上的PI3P调节的,与HIV Gag在血膜上的PI(4,5) P2的依赖不同.
- 了解Arc的PI3P依赖分泌途径,可以了解它在细胞间货物转移中的作用,并突出了保存和独特的病毒囊贩运机制.
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