Studying Y14-RNA interaction and co-phase separation

Abstract

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Y14 is a multifunctional RNA processing factor that plays an essential role in RNA metabolism and also participates in non-homologous end joining (NHEJ)-mediated repair of DNA double-strand breaks (DSBs). Our recent study has uncovered a long non-coding RNA (lncRNA) HOTAIRM1 that mediates the interaction between Y14 and the NHEJ factors. In this study, I first examined the molecular interaction between Y14 and RNA, in particular, HOTAIRM1. RNA electrophoretic mobility shift assay (REMSA) revealed that Y14 bound to multiple regions of HOTAIRM1. Our recent study also revealed that Y14 has the capability to undergo liquid-liquid phase separation (LLPS), and can co-phase-separate with HOTAIRM1. Therefore, I evaluated whether RNA structure or modification affects the LLPS properties of Y14. Moreover, since DNA damage induced m6A modification of HOTAIRM1, I also evaluated whether RNA methylation also affects the LLPS of Y14. My apparent data revealed that methylation enhanced Y14 LLPS. Since LLPS may involve electrostatic interactions of intrinsically disordered domains, I evaluated whether specific charge patterns can modulate a protein’s LLPS capacity. The multifunctionality of Y14 and its biochemical properties make it a valuable target for further research in understanding its role in RNA metabolism and DSB repair.