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Structure-function correlations of the insulin-linked polymorphic region

P Catasti1, X Chen, R K Moyzis

  • 1Theoretical Biology and Biophysics, Los Alamos National Laboratory, NM 87545, USA.

Journal of Molecular Biology
|December 6, 1996
PubMed
Summary
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The insulin minisatellite (ILPR) forms a hairpin G-quartet structure, influencing human insulin gene transcription and susceptibility to insulin-dependent diabetes mellitus (IDDM). This structural insight explains ILPR length polymorphism and its link to IDDM.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biophysics

Background:

  • The insulin-linked polymorphic region (ILPR) upstream of the human insulin gene is associated with insulin-dependent diabetes mellitus (IDDM).
  • ILPR length polymorphism correlates with insulin gene transcriptional activity and IDDM susceptibility.
  • The structural basis for ILPR polymorphism and its regulatory role remains unclear.

Purpose of the Study:

  • To elucidate the structural role of the ILPR in length polymorphism and transcriptional regulation.
  • To investigate the G-rich strand's secondary structure formation within the ILPR.

Main Methods:

  • Gel electrophoresis, circular dichroism (CD), and 1D/2D nuclear magnetic resonance (NMR) spectroscopy were employed.
  • Structural analysis of synthetic ILPR DNA sequences (d(G4TGTG4) and d(G4TGTG4ACAG4TGTG4)) was performed.

Related Experiment Videos

  • In vitro replication assays were used to assess structural formation in longer ILPR repeats.

    • Main Results:

    • The G-rich strand of the ILPR adopts an intramolecular hairpin G-quartet structure.
    • Detailed NMR analysis defined the folding, G-tetrad stacking, and loop interactions within the hairpin structure.
    • Multiple hairpin G-quartet structures were observed in longer ILPR repeats, potentially explaining replication slippage and length polymorphism.
    • Mutations destabilizing the hairpin G-quartet structure reduced insulin gene transcriptional activity.

    Conclusions:

    • The hairpin G-quartet structure is a key determinant of ILPR length polymorphism.
    • This G-quartet structure directly correlates with and influences the transcriptional activity of the human insulin gene.
    • The findings provide a structural mechanism linking ILPR variation to IDDM susceptibility.