Integrative MeRIP-Seq and RNA-Seq Analyses Reveal Innate Immune and Infection-Related Transcriptomic Changes upon...

Area of Science:

• Molecular Biology
• Epigenetics
• Immunology

Background:

• Methyltransferase-like 3 (METTL3) is a key enzyme catalyzing N6-methyladenosine (m⁶A) mRNA modifications.
• METTL3 influences RNA splicing, stability, and distribution, playing a crucial role in gene regulation.
• The global impact of METTL3 on mRNA methylation patterns remains incompletely understood.

Purpose of the Study:

• To elucidate the global landscape of mRNA methylation alterations driven by METTL3.
• To identify specific genes and pathways regulated by METTL3-mediated m⁶A modification.
• To explore the role of METTL3 in innate immune responses against pathogens.

Main Methods:

• Constructed a HEK293T cell line with METTL3 depletion.
• Performed RNA sequencing (RNA-seq) and methylated RNA Immunoprecipitation Sequencing (MeRIP-seq).
• Utilized quantitative Reverse Transcription PCR (qRT-PCR) for gene expression validation.

Main Results:

• Characterized the mRNA methylation alteration landscape upon METTL3 depletion.
• Identified 5763 hypomethylated and 269 hypermethylated genes after METTL3 silencing.
• Discovered METTL3-regulated innate immune genes (e.g., MYD88, RIG-1, CYLD, IRF9) enriched in pathogen infection and immune response pathways.

Conclusions:

• METTL3 significantly impacts the global mRNA methylation profile.
• METTL3-regulated genes are involved in crucial innate immune pathways, including responses to Shigellosis, Yersinia, and HIV-1.
• METTL3 and its regulated genes represent potential therapeutic targets for combating infections.