The Proteins Interacting with Prmt5 in Medaka (Oryzias latipes) Identified by Yeast Two-Hybridization

Abstract

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.

Maternal methylosome protein 50 is essential for embryonic development in medaka Oryzias latipes

“…It was first discovered as a close associate of the protein arginine methyltransferase 5 (Prmt5) in the methylosome complex (Friesen et al, 2002). Both Prmt5 and Mep50 are conserved in animals from fish to humans Homo sapiens (Shen et al, 2020). Mep50 can significantly enhance the methyltransferase activity of Prmt5 and is necessary for Prmt5 to methylate the histones H2A and H4 through substrate recognition and orientation (Ho et al, 2013).…”

“…Both Prmt5 and Mep50 are conserved in animals from fish to humans Homo sapiens (Shen et al, 2020). Mep50 can significantly enhance the methyltransferase activity of Prmt5 and is necessary for Prmt5 to methylate the histones H2A and H4 through substrate recognition and orientation (Ho et al, 2013).…”