Germline Maintenance Through the Multifaceted Activities of GLH/Vasa in Caenorhabditis elegans P Granules
| Autor: | Dustin L. Updike, Sarah E. Holbrook, J. Heath Fuqua, Jesse D. Rochester, Catherine S. Sharp, Emily L. Xu, Elisabeth A. Marnik |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Developmental and Behavioral Genetics
Small RNA Proteasome Endopeptidase Complex GLH-1 Eukaryotic Initiation Factor-3 Protein domain Glycine Piwi-interacting RNA Investigations germline Cytoplasmic Granules Ribosome DEAD-box RNA Helicases 03 medical and health sciences P granules 0302 clinical medicine Protein Domains DDX4 Genetics CRISPR Vasa Animals Caenorhabditis elegans Caenorhabditis elegans Proteins 030304 developmental biology 0303 health sciences biology Chemistry urogenital system COP9 Signalosome Complex Wild type RNA Helicase Translation (biology) PCI complex Argonaute biology.organism_classification 3. Good health Cell biology Germ Cells Mutation biology.protein C. elegans germ granules 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Genetics |
| ISSN: | 1943-2631 0016-6731 |
| Popis: | Vasa homologs are ATP-dependent DEAD-box helicases, multipotency factors, and critical components that specify and protect the germline. They regulate translation, amplify piRNAs, and act as RNA solvents; but, the limited availability of mutagenesis-derived alleles and their wide... Vasa homologs are ATP-dependent DEAD-box helicases, multipotency factors, and critical components that specify and protect the germline. They regulate translation, amplify piwi-interacting RNAs (piRNAs), and act as RNA solvents; however, the limited availability of mutagenesis-derived alleles and their wide range of phenotypes have complicated their analysis. Now, with clustered regularly interspaced short palindromic repeats (CRISPR/Cas9), these limitations can be mitigated to determine why protein domains have been lost or retained throughout evolution. Here, we define the functional motifs of GLH-1/Vasa in Caenorhabditis elegans using 28 endogenous, mutant alleles. We show that GLH-1’s helicase activity is required to retain its association with P granules. GLH-1 remains in P granules when changes are made outside of the helicase and flanking domains, but fertility is still compromised. Removal of the glycine-rich repeats from GLH proteins progressively diminishes P-granule wetting-like interactions at the nuclear periphery. Mass spectrometry of GLH-1-associated proteins implies conservation of a transient piRNA-amplifying complex, and reveals a novel affinity between GLH-1 and three structurally conserved PCI (26S Proteasome Lid, COP9, and eIF3) complexes or “zomes,” along with a reciprocal aversion for assembled ribosomes and the 26S proteasome. These results suggest that P granules compartmentalize the cytoplasm to exclude large protein assemblies, effectively shielding associated transcripts from translation and associated proteins from turnover. Within germ granules, Vasa homologs may act as solvents, ensuring mRNA accessibility by small RNA surveillance and amplification pathways, and facilitating mRNA export through germ granules to initiate translation. |
| Databáze: | OpenAIRE |
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