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Peer-reviewedCC BY 4.0International Journal of Molecular Sciences

Regulatory Elements and Splicing Factors Govern Mutually Exclusive Splicing of DmRdl Exon 3

  • Zijiao Song1
  • Yichi Zhang1
  • Kaixiang Lin1
  • Xiugen Wang1
  • Yanan Duan1
  • Jian Bao1
  • Chris Bass2
  • Chunqing Zhao1
  1. 1State Key Laboratory of Agricultural and Forestry Biosecurity, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
  2. 2Centre for Ecology and Conservation, University of Exeter, Penryn Campus, Penryn, Cornwall TR10 9FE, UK
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Abstract

Ionotropic γ-aminobutyric acid (GABA) receptors are major inhibitory receptors in insect central nervous systems, with their diversity partly generated by alternative splicing. In Drosophila melanogaster, mutually exclusive alternative splicing (MEAS) of the resistant to dieldrin gene (Rdl) exon 3 (E3) produces Rdl-3a and Rdl-3b, but its regulatory mechanism remains unclear. In the present study, two intronic regulatory elements (IREs) and one exonic regulatory element (ERE) were identified in the exon 2 (E2)–exon 4 (E4) region of DmRdl precursor mRNA (pre-mRNA). The IRE1 mutation decreased exon 3b (E3b) inclusion from 54.12% to 3.95%, whereas the IRE2 mutation increased E3b inclusion from 61.70% to 97.82%. The ERE mutation significantly increased E3b inclusion both in vitro and in vivo, while the IRE mutations were lethal in vivo. The results of RNA pull-down, mass spectrometry, and RNAi demonstrated that the splicing factors, CG30122 and Rbp1-like bind to the ERE and exert opposite effects on E3b inclusion. The results of STRING network analysis supported the functional interaction between CG30122 and Rbp1-like, while molecular docking provided structural support for their potential interaction with the ERE-containing RNA sequence. This study reveals the molecular regulatory network underlying the MEAS of DmRdl E3, and and provides a foundation for investigating other MEAS exons, such as E6 and E9 in insect Rdl.

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Cite

BibTeXRIS
BibTeX
@article{song2026regulatory,
  title = {{Regulatory Elements and Splicing Factors Govern Mutually Exclusive Splicing of DmRdl Exon 3}},
  author = {Zijiao Song and Yichi Zhang and Kaixiang Lin and Xiugen Wang and Yanan Duan and Jian Bao and Chris Bass and Chunqing Zhao},
  journal = {International Journal of Molecular Sciences},
  year = {2026},
  month = oct,
  doi = {10.3390/ijms27219493},
  url = {https://www.mdpi.com/1422-0067/27/21/9493},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Regulatory Elements and Splicing Factors Govern Mutually Exclusive Splicing of DmRdl Exon 3
AU  - Zijiao Song
AU  - Yichi Zhang
AU  - Kaixiang Lin
AU  - Xiugen Wang
AU  - Yanan Duan
AU  - Jian Bao
AU  - Chris Bass
AU  - Chunqing Zhao
PY  - 2026
DA  - 2026/10/24
JO  - International Journal of Molecular Sciences
DO  - 10.3390/ijms27219493
UR  - https://www.mdpi.com/1422-0067/27/21/9493
AB  - Ionotropic γ-aminobutyric acid (GABA) receptors are major inhibitory receptors in insect central nervous systems, with their diversity partly generated by alternative splicing. In Drosophila melanogaster, mutually exclusive alternative splicing (MEAS) of the resistant to dieldrin gene (Rdl) exon 3 (E3) produces Rdl-3a and Rdl-3b, but its regulatory mechanism remains unclear. In the present study, two intronic regulatory elements (IREs) and one exonic regulatory element (ERE) were identified in the exon 2 (E2)–exon 4 (E4) region of DmRdl precursor mRNA (pre-mRNA). The IRE1 mutation decreased exon 3b (E3b) inclusion from 54.12% to 3.95%, whereas the IRE2 mutation increased E3b inclusion from 61.70% to 97.82%. The ERE mutation significantly increased E3b inclusion both in vitro and in vivo, while the IRE mutations were lethal in vivo. The results of RNA pull-down, mass spectrometry, and RNAi demonstrated that the splicing factors, CG30122 and Rbp1-like bind to the ERE and exert opposite effects on E3b inclusion. The results of STRING network analysis supported the functional interaction between CG30122 and Rbp1-like, while molecular docking provided structural support for their potential interaction with the ERE-containing RNA sequence. This study reveals the molecular regulatory network underlying the MEAS of DmRdl E3, and and provides a foundation for investigating other MEAS exons, such as E6 and E9 in insect Rdl.
SN  - 1422-0067
ER  -