Direct and heterologous approaches to identify the LET-756/FGF interactome

Area of Science:

• Molecular Biology
• Developmental Biology
• Cell Biology

Background:

• Fibroblast growth factors (FGFs) are key signaling proteins regulating cell functions.
• LET-756 is a C. elegans FGF essential for development, with nuclear and secreted localization.
• The nuclear roles and interactions of LET-756 are not fully understood.

Purpose of the Study:

• To identify nuclear factors interacting with LET-756.
• To establish an interaction map for LET-756 and its binding partners.
• To explore conserved protein-protein interactions across species.

Main Methods:

• Yeast two-hybrid screening using LET-756 as bait with C. elegans and human cDNA libraries.
• Validation of interactions via co-immunoprecipitation in mammalian cells.
• Identification of conserved orthologs and assessment of binary interactions.

Main Results:

• Identified nuclear interactors including PAL-1 (transcription regulator) and RPS-16 (ribosomal component).
• Discovered KIN-3 and KIN-10 (orthologs of CK2alpha/beta) as novel LET-756 partners.
• Confirmed interaction with 14-3-3beta and its C. elegans orthologs FTT-1 and FTT-2/PAR-5.

Conclusions:

• This study provides the first comprehensive interaction map for LET-756.
• Identified interactors are involved in critical developmental and biochemical processes.
• The findings contribute to understanding FGF signaling and protein interactions across species.