PXD072041

PXD072041 is an original dataset announced via ProteomeXchange.

Title	An Evolutionarily Conserved N-terminal Domain of RRF-3 Governs GTSF-1 Binding Specificity in Nematodes
Description	GTSF1 is an essential activating cofactor for PIWI proteins in many metazoans. In the nematode Caenorhabditis elegans, however, GTSF-1 does not bind PIWI, but is associated with the RNA-dependent RNA polymerase RRF-3, supporting endo-siRNA (26G-RNA) biogenesis. Here, we demonstrate that this rewiring is deeply conserved across nematodes. For C. briggsae and Pristionchus pacificus we show that GTSF-1 interacts with RRF-3 and is essential for 26G-RNA production and fertility. We map this interaction to an N-terminal domain of RRF-3, termed the GTSF-1 interacting domain (GID), and show that the GTSF-1 zinc finger region alone is sufficient for binding. Mutagenesis identifies critical residues mediating this interaction and reveals that GTSF-1 stability depends on RRF-3. Other RdRPs possess GID-like domains, which we propose to bind GTSF-1-related proteins. Phylogenomic and structural analyses support GTSF-1–RRF-3 interactions across all major nematode lineages and map the shift in GTSF-1 activity to the last common nematode ancestor. We propose that binding of GTSF-1 induces conformational changes in RRF-3 that facilitate RdRP complex assembly and activate its function, paralleling its role as a PIWI activator.
HostingRepository	PRIDE
AnnounceDate	2026-08-20
AnnouncementXML	Submission_2026-08-19_22:38:28.801.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	F Butter
SpeciesList	scientific name: Pristionchus pacificus; NCBI TaxID: NEWT:54126; scientific name: Caenorhabditis briggsae; NCBI TaxID: NEWT:6238;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2025-12-16 04:23:20	ID requested
⏵ 1	2026-08-19 22:38:29	announced

Dataset with its publication pending

submitter keyword: LC-MS/MS

Falk Butter
contact affiliation	Institute of Molecular Biology, Mainz, Germany
contact email	f.butter@imb.de
lab head
F Butter
contact affiliation	Quantitative Proteomics Institute of Molecular Biology (IMB)
contact email	f.butter@imb-mainz.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD072041
     1. Label: PRIDE project
     2. Name: An Evolutionarily Conserved N-terminal Domain of RRF-3 Governs GTSF-1 Binding Specificity in Nematodes