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The Proteins Interacting with Prmt5 in Medaka (Oryzias latipes) Identified by Yeast Two-Hybridization
Hao Shen1, Xiaosha Zhang1, Md Abdullah Al Hafiz1
1Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan 430079, China.
Background:
Prmt5 plays major role in regulation of gene expression, RNA processing, cell growth and differentiation, signal transduction, germ cell development, etc., in mammals. Prmt5 is also related to cancer. Knowing the proteins interacting with Prmt5 is important to understand Prmt5's function in cells. Although there have been reports on proteins binding with Prmt5 in mammals, the partner proteins of Prmt5 in fish are still unclear.
Objectives:
The objective was to obtain proteins that bind with Prmt5 in medaka, a model fish.
Methods:
Yeast two hybridization was adopted to achieve the objective. Medaka Prmt5 was used as a bait to fish the prey, binding proteins in a cDNA library of medaka. Co-immunoprecipitation and in silicon analysis were performed to study the interaction of medaka Mep50 and Prmt5.
Results:
Eight proteins were identified to bind with Prmt5 from 69 preliminary positive colonies. The binding proteins are methylosome protein 50 (Mep50), apolipoprotein A-I-like (Apo-AI), PR domain containing protein 1a with zinc fingers (Prdm1a), Prdm1b, T-cell immunoglobulin mucin family member 3 (Tim-3), phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazolesuccinocarboxamide synthase (Paics), NADH dehydrogenase subunit 4 (ND4) and sciellin (Scl). Co-immunoprecipitation confirmed the interaction of medaka Prmt5 and Mep50. Predicted structures of medaka Prtm5 and Mep50 are similar to that of human PRMT5 and MEP50.
Conclusion:
Medaka Mep50, Prdm1a, Prdm1b, Apo-AI, Tim-3, Paics, ND4, and Scl bind with Prmt5.
Insights
Protein arginine methyltransferase 5 (Prmt5) interacts with eight proteins in medaka fish, including methylosome protein 50 (Mep50). This study identifies novel Prmt5 binding partners in fish, crucial for understanding its cellular functions.
Area of Science:
- Molecular Biology
- Proteomics
- Fish Model Organisms
Background:
- Protein arginine methyltransferase 5 (Prmt5) is vital for gene regulation, cell growth, and development in mammals.
- Prmt5's role in cancer highlights the importance of understanding its interactions.
- While Prmt5 interactions are known in mammals, they remain uncharacterized in fish.
Purpose of the Study:
- To identify proteins that bind with Prmt5 in the medaka fish model.
- To expand the understanding of Prmt5's functional network in a vertebrate system.
Main Methods:
- Yeast two-hybrid screening using medaka Prmt5 as bait.
- Co-immunoprecipitation to validate protein interactions.
- In silico analysis to compare structural similarities.
Main Results:
- Eight proteins were identified as Prmt5 binding partners in medaka.
- Key interactors include methylosome protein 50 (Mep50), PR domain containing proteins 1a and 1b (Prdm1a, Prdm1b), and apolipoprotein A-I-like (Apo-AI).
- Co-immunoprecipitation confirmed the Prmt5-Mep50 interaction; structural predictions showed similarity to human counterparts.
Conclusions:
- Medaka Prmt5 interacts with Mep50, Prdm1a, Prdm1b, Apo-AI, T-cell immunoglobulin and mucin-domain containing-3 (Tim-3), phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthase (Paics), NADH dehydrogenase subunit 4 (ND4), and sciellin (Scl).
- These findings establish a foundational understanding of Prmt5's protein interactome in fish.
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