Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans

eLife

Hannah L Hertz¹  Wen Tang¹  Ian F Price²  Benjamin Pastore²  Jillian Wagner³ 

[1] Department of Biological Chemistry and Pharmacology, Columbus, United States;Center for RNA Biology, Columbus, United States;Department of Biological Chemistry and Pharmacology, Columbus, United States;Center for RNA Biology, Columbus, United States;Ohio State Biochemistry Program, Columbus, United States;Department of Biological Chemistry and Pharmacology, Columbus, United States;Department of Molecular Genetics, Ohio State University, Columbus, United States;

关键词: germ granules;    LOTUS domain protein;    proximity labeling;    p granules;    vasa proteins;    C. elegans;   

DOI  :  10.7554/eLife.72276

来源: eLife Sciences Publications, Ltd

【 摘 要 】

The germ line produces gametes that transmit genetic and epigenetic information to the next generation. Maintenance of germ cells and development of gametes require germ granules—well-conserved membraneless and RNA-rich organelles. The composition of germ granules is elusive owing to their dynamic nature and their exclusive expression in the germ line. Using Caenorhabditis elegans germ granule, called P granule, as a model system, we employed a proximity-based labeling method in combination with mass spectrometry to comprehensively define its protein components. This set of experiments identified over 200 proteins, many of which contain intrinsically disordered regions (IDRs). An RNA interference-based screen identified factors that are essential for P granule assembly, notably EGGD-1 and EGGD-2, two putative LOTUS-domain proteins. Loss of eggd-1 and eggd-2 results in separation of P granules from the nuclear envelope, germline atrophy, and reduced fertility. We show that IDRs of EGGD-1 are required to anchor EGGD-1 to the nuclear periphery while its LOTUS domains are required to promote the perinuclear localization of P granules. Taken together, our work expands the repertoire of P granule constituents and provides new insights into the role of LOTUS-domain proteins in germ granule organization.

【 授权许可】

CC BY