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WAGO-4 Localization to Germ Granules Is Important for Its Accurate Small RNA Loading

ABSTRACT: There were several MS experiments used in this study: 1) Proximity labeling of the P granules 2) Protein levels estimation from whole worm lysates and 3) wago-4 KO expression check (PRM analysis). 1) Proximity labeling strategy was used to estimate P granule composition in 3xFG mutant (FG domains of GLH-1, GLH-2, and GLH-4 mutated) and compare it to the WT. PGL-3 was tagged with TurboID at its native locus using CRISPR-Cas9. Biotinylation level was determined after streptavidin pull-down followed by MS analysis, serving as a proxy for protein enrichment in P granules. 2) Whole-worm lysate MS analysis was performed on 3xFG mutant worms to assess the worm proteome at an elevated temperature (26 °C), where these mutants exhibit a fecundity phenotype. 3) A STOP codon was introduced in the second exon of the WAGO-4 gene at its native locus in C. elegans (WAGO-4 KO). To assess whether the WAGO-4 protein is expressed in this strain, lysates from WAGO-4 KO (in triplicate) and N2 (WT control) were analyzed using PRM-MS.

INSTRUMENT(S):

ORGANISM(S): Caenorhabditis Elegans

TISSUE(S): Whole Body

SUBMITTER: Richard Imre

LAB HEAD: Stela Jelenic

PROVIDER: PXD062557 | Pride | 2025-10-14

REPOSITORIES: Pride

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			Action	DRS
	20220512_E3_RSLC1_Jelenic_Saha_IMBA_ID722_onbead_dig_5per_4x3samples_celegans_Step2_Limma_updatedPOIdb_quanOnAll_ratioInverted.pdResult	Other
	20220512_E3_RSLC1_Jelenic_Saha_IMBA_ID722_onbead_dig_5per_4x3samples_celegans_Step2_Limma_updatedPOIdb_uniqueRazor_SumNorm_ratioInverted.pdResult	Other
	20220512_E3_RSLC1_Jelenic_Saha_IMBA_ID722_onbead_dig_5per_control_1A_N2_A.raw	Raw
	20220512_E3_RSLC1_Jelenic_Saha_IMBA_ID722_onbead_dig_5per_control_1B_N2_B.raw	Raw
	20220512_E3_RSLC1_Jelenic_Saha_IMBA_ID722_onbead_dig_5per_control_1C_N2_C.raw	Raw

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