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Luige, J.

Publications and source records attributed to Luige, J..

2 recordsLinked to original sources

Fine-tuning m6A and METTL3 levels have profound impact on cellular proliferation and protein synthesis

The RNA modification m6A is the most abundant internal RNA modification in eukaryotic mRNAs and long non-coding RNAs and has been implicated in diverse and important biological processes. Notably, m6A has been associated with both pro and anti-tumorigenic roles depending on cellular and biological context. In basal-like triple negative breast cancer (TNBC), heterozygous loss of the m6A methyltransferase METTL3 and increased levels of the m6A demethylase FTO are associated with poor prognosis and an increased risk of metastasis. Here, using CRISPR generated METTL3 heterozygous knockout TNBC cell lines (MDA-MB-468) and Nanopore direct RNA sequencing, we characterise transcriptome-wide changes in m6A modification patterns following partial loss of METTL3. We reveal that partial loss of global m6A is associated with preferential changes in the methylation status of transcripts involved in translational control, leading to an increase in translational output and proliferative capacity. In contrast, strong pharmacologic inhibition of METTL3 suppresses translation and proliferation. Our findings highlight how m6A levels differentially regulate gene expression in a dose dependent manner and provides a deeper molecular understanding of the RNA modification m6A and its role in fine-tuning translation and affecting both tumorigenesis and cancer progression.

molecular biology↗

Identification and prediction of G-quadruplex RNA-binding proteins with roles in transcription and phase separation

RNA-binding proteins are central for many biological processes and their large-scale identification has demonstrated a broad range of functions. RNA G-quadruplexes are important regulatory elements occurring in both coding and non-coding transcripts, yet our knowledge of their structure-based interactions is limited. Here starting from theoretical prediictions, we show experimentally that a large number of chromatin-binding proteins bind to RNA G-quadruplexes and we classify these based on their RNA G-quadruplex-binding potential. Combining experimental identification of nuclear RNA G-quadruplex-binding proteins with computational analysis, we create a prediction tool that can assign probability score for a protein that it binds RNA G-quadruplexes. We show that predicted G-quadruplex RNA-binding proteins exhibit high degree of protein disorder and hydrophilicity, and suggest involvement in both transcription and phase-separation into membrane-less organelles, particularly the nucleolus. Finally, we present this tool as a web application for estimating RNA G4-binding propensity for proteins of interest at http://service.tartaglialab.com/new_submission/clever_G4_classifier.

molecular biology↗