Abstract
In this study, the transcriptional regulation of PI3KC3 by three transcription factors (PPARγ, PPARα, and STAT3) and the potential role of PI3KC3 in mediating lipid accumulation were determined in yellow catfish Pelteobagrus fulvidraco. The 5’-deletion assay, overexpression assay, site-mutation assay, and electrophoretic mobility shift assay suggested that PPARα, PPARγ, and STAT3 negatively regulated the promoter activity of pi3kc3. Moreover, the transcriptional inactivation of pi3kc3 was directly mediated by PPARα and PPARγ under fatty acid (FA) treatment. Using primary hepatocytes from yellow catfish, FA incubation significantly increased triacylglyceride (TG) content, non-esterified fatty acid (NEFA) content, and lipid drops (LDs) content, the mRNA level of pparα, pparγ, stat3, and dnmt3b, the protein level of PPARα, PPARγ, and STAT3, and the methylation level of pi3kc3, but significantly reduced the mRNA and protein level of PI3KC3. Our findings offer new insights into the mechanisms for transcriptional regulation of PI3KC3 and for PI3KC3-mediated lipid accumulation in fish.
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Abbreviations
- FA:
-
Fatty acid
- GAPDH:
-
Glyceraldehyde-3-phosphate dehydrogenase
- HPRT:
-
Hypoxanthineguanine phosphoribosyltransferase
- LDs:
-
Lipid drops
- NEFA:
-
Non-esterified fatty acid
- Mut:
-
Mutagenesis
- NP:
-
Nuclear protein
- PPAR:
-
Peroxisome proliferator-activated receptor
- OE:
-
Overexpression
- PI3K:
-
Phosphatidylinositol-3 kinase
- RPL7:
-
Ribosomal protein L7
- STAT3:
-
Signal transducers and activators of transcription proteins 3
- TG:
-
Triacylglyceride
- TUBA:
-
Tubulin alpha chain
- UBCE:
-
Ubiquitin-conjugating enzyme
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This work was supported by the National Natural Science Foundation of China (Grant No. 31902381).
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MQ Zhuo designed the experiments; MQ Zhuo and J Chen performed the experiments, and analyzed the samples with the help of ML Wu and WB Wang; J Chen and ML Wu analyzed the data; and J Chen and MQ Zhuo wrote the manuscript. MQ Zhuo revised the manuscript. All the authors approved the manuscript.
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Zhuo, MQ., Chen, J., Wu, ML. et al. Novel insights for PI3KC3 in mediating lipid accumulation in yellow catfish Pelteobagrus fulvidraco. Fish Physiol Biochem 48, 571–583 (2022). https://doi.org/10.1007/s10695-022-01071-7
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DOI: https://doi.org/10.1007/s10695-022-01071-7