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METTL3 activates PERK-eIF2α dependent coelomocyte apoptosis by targeting the endoplasmic reticulum

Dongdong Li1, Ming Guo1, Zhimeng Lv1

  • 1State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Ningbo University, PR China.

Biochimica Et Biophysica Acta. Gene Regulatory Mechanisms
|March 18, 2023
PubMed

Insights

Invertebrate methyltransferase-like 3 (METTL3) in Apostichopus japonicus promotes Vibrio splendidus-induced coelomocyte apoptosis. AjMETTL3 regulates the endoplasmic reticulum-related degradation pathway by targeting AjSEL1L mRNA, impacting immune responses.

Area of Science:

  • Immunology
  • Molecular Biology
  • Marine Biology

Background:

  • N6-methyladenosine (m6A) modification is crucial in vertebrate physiological and disease processes.
  • Methyltransferase-like 3 (METTL3) is a key enzyme in m6A methylation, but its role in invertebrates remains largely unexplored.
  • The sea cucumber Apostichopus japonicus serves as a model organism to investigate invertebrate immune mechanisms.

Purpose of the Study:

  • To elucidate the function of Apostichopus japonicus METTL3 (AjMETTL3) in the innate immune response against Vibrio splendidus infection.
  • To investigate the molecular mechanism underlying AjMETTL3-mediated regulation of coelomocyte apoptosis.
  • To identify potential targets and pathways regulated by AjMETTL3 in coelomocytes.

Main Methods:

  • Gene expression analysis (qRT-PCR) to assess AjMETTL3 induction.
  • Coelomocyte culture and manipulation (overexpression, silencing) to study apoptosis.
  • m6A-sequencing (m6A-seq) to identify m6A-modified transcripts.
  • Western blotting to analyze protein levels and pathway activation.
  • Functional assays to confirm AjSEL1L's role in apoptosis.

Main Results:

  • AjMETTL3 expression and m6A levels were significantly upregulated in coelomocytes upon Vibrio splendidus challenge.
  • AjMETTL3 overexpression promoted, while silencing inhibited, V. splendidus-induced coelomocyte apoptosis.
  • m6A-seq identified the endoplasmic reticulum-associated degradation (ERAD) pathway as enriched, with suppressor/enhancer of Lin-12-like (AjSEL1L) as a direct target of AjMETTL3.
  • AjMETTL3 reduced AjSEL1L mRNA stability, leading to decreased AjSEL1L levels, increased AjOS9 and Ajp97 transcription, ER stress, and subsequent coelomocyte apoptosis via the PERK-eIF2α pathway.

Conclusions:

  • Invertebrate METTL3 plays a critical role in regulating apoptosis during immune challenges.
  • AjMETTL3 mediates coelomocyte apoptosis by targeting AjSEL1L within the ERAD pathway.
  • The AjMETTL3-AjSEL1L interaction influences the PERK-eIF2α signaling cascade, highlighting a conserved mechanism in invertebrate immunity.
Keywords:
ApoptosisMethyltransferase-like 3N6-methyladenosineProtein kinase RNA-like endoplasmic reticulum kinaseSuppressor/enhancer of Lin-12-like

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