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i-Motif, not G-quadruplex, stability regulates insulin expression.

Dilek Guneri1,2, Christopher J Morris1, Yiliang Ding3

  • 1School of Pharmacy, University College London, 29-39 Brunswick Square, London WC1N 1AX, United Kingdom.

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|January 29, 2026
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Summary
This summary is machine-generated.

Transcriptional activation of the human insulin gene requires both G-quadruplex and i-motif structures to form. Promoter activity correlates with i-motif stability, suggesting a dynamic interplay in gene regulation.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Biophysics

Background:

  • The insulin-linked polymorphic region (ILPR) is a variable tandem repeat in the human insulin gene promoter.
  • ILPR sequences can form G-quadruplexes (G4) and i-motifs (IM), DNA structures implicated in gene regulation.
  • The precise roles of G4 and IM in regulating insulin gene expression, especially when co-occurring, are not fully understood.

Purpose of the Study:

  • To investigate the relationship between ILPR sequence diversity, DNA structure formation (G4 and IM), and insulin gene regulation.
  • To determine the conditions under which G4 and IM structures influence transcriptional activity in response to glucose.
  • To elucidate the specific roles of G4 and IM structures in modulating insulin gene promoter activity.

Main Methods:

  • Characterization of nine ILPR-based DNA sequences using biophysical techniques.
  • Assessment of DNA structure stability and formation.
  • Luciferase reporter assays to measure promoter activity under varying glucose conditions.

Main Results:

  • Transcriptional activation of the insulin gene promoter was observed only when both G-quadruplex and i-motif structures could form.
  • Insulin gene promoter activity showed a positive correlation with i-motif stability, but not with G-quadruplex stability.
  • Different combinations of G4 and IM structures did not induce transcription, highlighting the necessity of both structures.

Conclusions:

  • A model is proposed where G-quadruplexes may initiate transcription, while i-motifs modulate insulin gene expression.
  • The formation and stability of both G-quadruplexes and i-motifs are crucial for glucose-responsive insulin gene regulation.
  • These DNA structures represent a dynamic, interdependent system with potential as therapeutic targets for gene expression modulation.