Chen Wenjun, Hu Yabing, Lang Charles F, Brown Jordan S, Schwabach Sierra, Song Xiaoyan, Zhang Ying, Munro Edwin, Bennett Karen, Zhang Donglei, Lee Heng-Chi
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China 430074.
Genetics. 2020 Jun;215(2):421-434. doi: 10.1534/genetics.120.303052. Epub 2020 Apr 3.
P granules are phase-separated liquid droplets that play important roles in the maintenance of germ cell fate in Both the localization and formation of P granules are highly dynamic, but mechanisms that regulate such processes remain poorly understood. Here, we show evidence that the VASA-like germline RNA helicase GLH-1 couples distinct steps of its ATPase hydrolysis cycle to control the formation and disassembly of P granules. In addition, we found that the phenylalanine-glycine-glycine repeats in GLH-1 promote its localization at the perinucleus. Proteomic analyses of the GLH-1 complex with a GLH-1 mutation that interferes with P granule disassembly revealed transient interactions of GLH-1 with several Argonautes and RNA-binding proteins. Finally, we found that defects in recruiting the P granule component PRG-1 to perinuclear foci in the adult germline correlate with the fertility defects observed in various GLH-1 mutants. Together, our results highlight the versatile roles of an RNA helicase in controlling the formation of liquid droplets in space and time.
P颗粒是相分离的液滴,在秀丽隐杆线虫生殖细胞命运维持中发挥重要作用。P颗粒的定位和形成都是高度动态的,但调节这些过程的机制仍知之甚少。在这里,我们证明了VASA样生殖系RNA解旋酶GLH-1将其ATPase水解循环的不同步骤耦合起来,以控制P颗粒的形成和分解。此外,我们发现GLH-1中的苯丙氨酸-甘氨酸-甘氨酸重复序列促进其在核周的定位。对具有干扰P颗粒分解的GLH-1突变的GLH-1复合物进行蛋白质组学分析,揭示了GLH-1与几种AGO蛋白和RNA结合蛋白的瞬时相互作用。最后,我们发现,在成年生殖系中将P颗粒成分PRG-1募集到核周焦点的缺陷与在各种GLH-1突变体中观察到的生育缺陷相关。总之,我们的结果突出了一种RNA解旋酶在时空上控制液滴形成中的多种作用。