PhD Scientific Days 2026

Molecular Medicine 3.

Flipping the Switch: An Inducible TurboID System for Mapping Interactome in C. elegans

Előadó neve

Khatatneh, Saba

Neptune code

EU8CCM

Előadó munkahelye

Semmelweis University, Institute of Biochemistry and Molecular Biology, Department of Molecular Biology

Előadó telefonszáma

+36704115915

Előadó e-mail címe

khatatneh.saba@phd.semmelweis.hu

Az előadás címe

Flipping the Switch: An Inducible TurboID System for Mapping Interactome in C. elegans

Szerző(k) neve és munkahelye

Saba Khatatneh¹, Milán Somogyvári¹, Csaba Sőti¹

1: Semmelweis University, Institute of Biochemistry and Molecular Biology, Department of Molecular Biology

Bemutatás módja

Szóbeli

Szekció

Molecular Medicine 3.

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Introduction: Mapping protein-protein interactions in vivo with temporal control remains a major challenge. Proximity labelling with TurboID offers rapid biotinylation of neighbouring proteins, but current implementations in C. elegans rely on constitutive overexpression, introducing developmental artefacts and masking dynamic interactions. A controllable system that activates labelling only at a chosen time point would enable cleaner, temporally resolved interactome studies.

Aims: To develop the first doxycycline-inducible TurboID proximity-labelling system in C. elegans and validate it using HSP-90, a conserved molecular chaperone with a wide range of interactors involved in all types of cellular processes, making it a great model as a bait protein. Besides, this paradigm gives us the chance to fill gaps in HSP90 function in the proliferating and terminally differentiated tissues of C. elegans.

Methods: A bipartite system was engineered: a constitutive pDriver (Pprl-28::rtTA(Q)/tTS) and an inducible pResponder (TRE::HSP-90::GFP::TurboID), integrated via biolistic transformation. A homozygous double-transgenic strain was established through crossing and multi-generational selection. Induction, expression kinetics, and biotinylation efficiency were characterised by fluorescence microscopy, qPCR and Western blotting. The HSP-90 inhibitor 17-DMAG is being evaluated as a pharmacological negative control using the heat shock response marker hsp-70 qPCR as a preliminary readout.

Results: The system showed tight regulation: no detectable expression in the absence of doxycycline and robust induction peaking at 24 hours post-treatment with 2 ng/µL doxycycline. Biotin supplementation (1 mM, 2 h) significantly enhanced labelling yield. Uninduced animals showed no increase in biotinylation even with exogenous biotin, confirming the absence of leak. Mass spectrometry-based interactome analysis is ongoing.

Conclusion: We established a tightly controlled, inducible proximity-labelling platform in C. elegans. By eliminating overexpression artefacts and enabling temporal control, this method is applicable to any bait protein and compatible with tissue-specific promoters for targeted interactome studies.

Funding: Stipendium Hungaricum Scholarship Programme, and OTKA PD 142838 research grant.

University

Semmelweis University

Supervisor

Milán Somogyvári and Csaba Sőti

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies after complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9745

Start

09:15

End

09:25