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Synthetic repeating sequence DNAs containing phosphorothioates: nuclease sensitivity and triplex formation.

L J Latimer1, K Hampel, J S Lee

  • 1Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

Nucleic Acids Research
|February 25, 1989
PubMed
Summary

Phosphorothioate modifications in DNA significantly lower melting temperatures (Tm) and dramatically impact DNA triplex formation. These changes suggest phosphorothioate groups induce subtle conformational alterations, potentially revealing new DNA structural variations.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • DNA structure and stability are crucial for biological processes.
  • Phosphorothioate modifications are used to alter DNA properties.

Purpose of the Study:

  • To investigate the effects of phosphorothioate modifications on DNA thermal stability and triplex formation.
  • To explore the relationship between phosphorothioate content and DNAase I resistance.

Main Methods:

  • Synthesis of over twenty repeating sequence DNAs containing phosphorothioates using DNA polymerase I.
  • Measurement of melting temperatures (Tm) of modified and parent DNA polymers.
  • Assessment of triplex formation under varying conditions.
  • Evaluation of resistance to pancreatic DNAse I digestion.

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Main Results:

  • All modified DNAs exhibited lower Tms compared to parent polymers, with greater decreases when the phosphorothioate was adjacent to a pyrimidine.
  • Phosphorothioate groups significantly affected triplex formation, with some sequences dismutating and others showing inhibited triplex formation.
  • Poly(dsG).poly(dC) showed an unusual decrease in Tm with increasing ionic strength.
  • DNAse I resistance did not correlate with phosphorothioate content, with some modified DNAs being resistant and others rapidly degraded.

Conclusions:

  • Phosphorothioate modifications induce small conformational changes in DNA.
  • These modifications may lead to the discovery of novel DNA conformational polymorphisms.
  • The impact of phosphorothioate groups on DNA structure is sequence-dependent and affects stability and interactions.