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MiR-210 regulates coelomocyte proliferation through targeting E2F3 in Apostichopus japonicus
Yi Zhang1, Yina Shao1, Zhimeng Lv1
1State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, PR China.
Abstract:
MiR-210 plays a crucial role in cell survival, migration, and regeneration in vertebrates. In our previous work, the expression of miR-210 was considerably induced in diseased Apostichopus japonicus with skin ulcer syndrome (SUS). To further explore the mechanism of miR-210 in regulating the SUS, this study identified E2F transcription factor 3 (E2F3), a candidate target of miR-210, from the sea cucumber A. japonicus via RNA-seq and RACE (designated as AjE2F3). A 1992 bp fragment representing the full-length cDNA of AjE2F3 was obtained, which includes an ORF of 1194 bp encoding a polypeptide of 398 amino acids with a molecular weight of 44.43 kDa. Expression profiling analysis suggested that the expression of AjE2F3 decreased while that of miR-210 increased in Vibrio splendidus-challenged sea cucumber coelomocytes. Dual-luciferase reporter assay revealed that miR-210 targeted AjE2F3 via binding to the 3'UTR region from 108 nt to 128 nt. MiR-210 overexpression in cultured coelomocytes repressed AjE2F3 at the mRNA level and reduced cell proliferation in vitro. Consistently, AjE2F3 overexpression significantly promoted coelomocyte proliferation, as assessed by MTT in vitro. Overall, our results indicated that miR-210 can suppress coelomocyte proliferation by targeting AjE2F3 in pathogen-challenged sea cucumbers.
Insights
MicroRNA-210 (miR-210) suppresses sea cucumber coelomocyte proliferation by targeting E2F transcription factor 3 (AjE2F3). This mechanism is crucial in pathogen-challenged sea cucumbers with skin ulcer syndrome.
Area of Science:
- Marine biology
- Molecular biology
- Immunology
Background:
- MicroRNA-210 (miR-210) is implicated in cell survival and regeneration.
- Previous studies showed miR-210 induction in diseased sea cucumbers (Apostichopus japonicus) with skin ulcer syndrome (SUS).
Purpose of the Study:
- To elucidate the mechanism of miR-210 in regulating SUS in A. japonicus.
- To identify and characterize a novel target of miR-210 involved in this process.
Main Methods:
- RNA sequencing (RNA-seq) and RACE were used to identify E2F transcription factor 3 (AjE2F3) as a miR-210 target.
- Dual-luciferase reporter assays confirmed miR-210 binding to the 3'UTR of AjE2F3.
- In vitro experiments assessed the effects of miR-210 and AjE2F3 overexpression on coelomocyte proliferation.
Main Results:
- AjE2F3 cDNA was identified, encoding a 398-amino acid polypeptide.
- miR-210 expression increased as AjE2F3 expression decreased in challenged coelomocytes.
- miR-210 overexpression repressed AjE2F3 and reduced coelomocyte proliferation, while AjE2F3 overexpression promoted it.
Conclusions:
- miR-210 suppresses coelomocyte proliferation by targeting AjE2F3 in pathogen-challenged sea cucumbers.
- This regulatory axis is a key factor in the sea cucumber's response to infection and disease.
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