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Updated: May 2, 2026
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
Published on: June 6, 2018
Identification of PLP2 and RAB5C as novel TPD52 binding partners through yeast two-hybrid screening
Hamideh Shahheydari1, Sarah Frost, Brian J Smith
1Children's Cancer Research Unit, Kids Research Institute, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia.
Abstract:
Tumor protein D52 (TPD52) is overexpressed in different cancers, but its molecular functions are poorly defined. A large, low-stringency yeast two-hybrid screen using full-length TPD52 bait identified known partners (TPD52, TPD52L1, TPD52L2, MAL2) and four other preys that reproducibly bound TPD52 and TPD52L1 baits (PLP2, RAB5C, GOLGA5, YIF1A). PLP2 and RAB5 interactions with TPD52 were confirmed in pull down assays, with interaction domain mapping experiments indicating that both proteins interact with a novel binding region of TPD52. This study provides insights into TPD52 functions, and ways to maximise the efficiency of low-stringency yeast two-hybrid screens.
Insights
This study identifies new binding partners for Tumor Protein D52 (TPD52), a protein overexpressed in cancers. These findings offer insights into TPD52
Area of Science:
- Molecular Biology
- Cancer Research
- Protein Interactions
Background:
- Tumor Protein D52 (TPD52) is frequently overexpressed in various cancers.
- The precise molecular functions of TPD52 remain largely uncharacterized.
- Understanding TPD52's interactions is crucial for elucidating its role in cancer.
Purpose of the Study:
- To identify novel protein interactors of TPD52.
- To investigate the functional implications of TPD52 interactions.
- To optimize yeast two-hybrid screening methodologies.
Main Methods:
- A large, low-stringency yeast two-hybrid screen was employed using TPD52 as bait.
- Interactions were validated using pull-down assays and domain mapping.
- TPD52 and TPD52-like 1 (TPD52L1) were used as bait proteins.
Main Results:
- The screen identified known TPD52 partners (TPD52, TPD52L1, TPD52L2, MAL2).
- Four novel interacting proteins (PLP2, RAB5C, GOLGA5, YIF1A) were reproducibly identified.
- PLP2 and RAB5C interactions with TPD52 were confirmed, with novel binding regions identified.
Conclusions:
- This research expands the known interactome of TPD52.
- The identified interactions provide new avenues for exploring TPD52's function in cancer.
- The study offers methodological insights for enhancing yeast two-hybrid screens.
