PXD014135

PXD014135 is an original dataset announced via ProteomeXchange.

Title	Germline maintenance through the multifaceted activities of GLH/Vasa in Caenorhabditis elegans P granules
Description	Vasa is a highly conserved member of the ATP-dependent DEAD box helicase family, a multipotency factor, and a critical component for the specification and maintenance of the germline. Its homologs have been shown to regulate translation, small RNA amplification, and serve as a molecular solvent for single-stranded RNA; however, the function of Vasa’s defining domains and what they interact with are unclear. To address this, 28 mutant alleles of the C. elegans Vasa homolog GLH-1 were generated in conserved motifs. Mutations in the flanking and helicase domains show that GLH-1 retains its association with P granules through its helicase activity and not through static interactions with other P-granule proteins. Changes outside of these domains retain GLH-1 in P granules but still compromise fertility, and removal of glycine-rich repeats progressively diminish P-granule wetting-like interactions at the nuclear periphery. A mutation that facilitates Vasa aggregation was previously leveraged in insects and mammals to identify the transient association of Vasa with piRNA amplifying Argonautes. This same mutation in GLH-1 also stimulates aggregation and association with Argonautes, suggesting that the transient amplifying complex is evolutionarily conserved even though the method of piRNA amplification in C. elegans is not. Mass spectrometry analysis of proteins that co-immunoprecipitate with wild type and mutant GLH-1 reveal an affinity for all three PCI (26S Proteasome Lid, COP9, eIF3) scaffolding complexes, which regulate protein turnover and translation, and an aversion for ribosomes and the 26S proteasome core. These results suggest that phase-separated P granules compartmentalize the cytoplasm to exclude large protein assemblies and emphasize the role of Vasa homologs in maintaining proteostasis.
HostingRepository	PRIDE
AnnounceDate	2019-11-08
AnnouncementXML	Submission_2019-11-08_00:43:05.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Dustin Updike
SpeciesList	scientific name: Caenorhabditis elegans; NCBI TaxID: 6239;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-06-05 01:55:39	ID requested
1	2019-06-06 23:34:47	announced
⏵ 2	2019-11-08 00:43:07	announced	2019-11-08: Updated publication reference for PubMed record(s): 31506335.

Marnik EA, Fuqua JH, Sharp CS, Rochester JD, Xu EL, Holbrook SE, Updike DL, P Granules. Genetics, 213(3):923-939(2019) [pubmed]

submitter keyword: GLH-1, Vasa, C. elegans, GLH-1 coIP

Dustin Lynn Updike
contact affiliation	The Mount Desert Island Biological Laboratory
contact email	dupdike@mdibl.org
lab head
Dustin Updike
contact affiliation	The MDI Biological Laboratory
contact email	dupdike@mdibl.org
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014135
     1. Label: PRIDE project
     2. Name: Germline maintenance through the multifaceted activities of GLH/Vasa in Caenorhabditis elegans P granules