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Hsa-miR-874-3p Reduces Endogenous Expression of RGS4-1 Isoform In Vitro.
Feng-Ling Xu1,2, Bao-Jie Wang1
1School of Forensic Medicine, China Medical University, Shenyang 110122, China.
Genes
|August 29, 2024
Summary
Reduced RGS4-1 levels in schizophrenia may be due to hsa-miR-874-3p. This microRNA targets the RGS4 gene
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Regulator of G-protein signaling 4-1 (RGS4-1) isoform levels are decreased in the dorsolateral prefrontal cortex (DLPFC) of individuals with schizophrenia.
- The underlying mechanisms for this reduction in RGS4-1 expression remain unclear.
- 3' untranslated regions (3'UTRs) are known regulators of mRNA splice variant levels.
Purpose of the Study:
- To investigate the regulatory role of the 3'UTR of the RGS4 gene in controlling RGS4-1 isoform expression.
- To identify specific microRNAs (miRNAs) that may target the RGS4 3'UTR and influence RGS4-1 levels.
- To elucidate the mechanism by which hsa-miR-874-3p affects RGS4-1 expression.
Main Methods:
- Construction of recombinant pmir-GLO vectors containing truncated RGS4 3' regulatory regions (3R1-3R6).
- Dual-luciferase reporter assays in HEK-293, SK-N-SH, and U87 cells to identify functional regions and predict miRNA binding sites.
- Western blot analysis and dual-luciferase assays following transient transfection with predicted miRNAs, including hsa-miR-874-3p mimics and mutated constructs.
Main Results:
- Functional regulatory regions within the RGS4 3'UTR were identified at positions +401-+789, +789-+1152, and +1562-+1990.
- Hsa-miR-874-3p, implicated in psychiatric disorders, was predicted to target the +789-+1152 region of the RGS4 3'UTR.
- Co-transfection with hsa-miR-874-3p mimic and the RGS4 3'UTR construct (3R1) decreased reporter activity, an effect reversed by a mutated construct (m3R1) lacking the +853-+859 region, indicating specific targeting. Hsa-miR-874-3p significantly reduced endogenous RGS4-1 expression.
Conclusions:
- Hsa-miR-874-3p directly inhibits the expression of the RGS4-1 isoform.
- The inhibitory mechanism involves hsa-miR-874-3p targeting of the specific sequence +853-+859 within the RGS4 3'UTR.
- This finding provides a potential molecular mechanism for reduced RGS4-1 levels observed in schizophrenia.
