The Dynamics of P Granule Liquid Droplets Are Regulated by the Caenorhabditis elegans Germline RNA Helicase GLH-1 via Its ATP Hydrolysis Cycle

• Published in: Genetics (Austin) Vol. 215; no. 2; pp. 421 - 434
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01-06-2020; Genetics Society of America
• Subjects: Adenosine triphosphatase; Adenosine Triphosphate - metabolism; Animals; Caenorhabditis elegans; Caenorhabditis elegans - genetics; Caenorhabditis elegans - growth & development; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Cell fate; CRISPR; Cytoplasmic Granules - metabolism; DEAD-box RNA Helicases - genetics; DEAD-box RNA Helicases - metabolism; Defects; Deoxyribonucleic acid; Dismantling; DNA; DNA helicase; Droplets; Fertility; Genetics; Glycine; Granular materials; Hydrolysis; Investigations; Liquid Crystals - chemistry; Localization; Mutation; Nematodes; Phenylalanine; Plasmids; Proteins; Proteomics; RNA helicase; RNA-binding protein; Young adults; VASA; phase separation; P granule; ATP hydrolysis; Germ granule; RNA helicase
• ISSN: 1943-2631; 0016-6731; 1943-2631
• DOI: 10.1534/genetics.120.303052

Abstract
Germ granules, also known as P granules in Caenorhabditis elegans, are phase-separated cellular bodies that are frequently found at the perinuclear region of germ cell nuclei in various animals. However...
P granules are phase-separated liquid droplets that play important roles in the maintenance of germ cell fate in Caenorhabditis elegans. 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.