Solution equilibria of the i-motif-forming region upstream of the B-cell lymphoma-2 P1 promoter

Biochimie. 2007 Dec;89(12):1562-72. doi: 10.1016/j.biochi.2007.07.026. Epub 2007 Aug 6.

Abstract

The 5'-end of the P1 promoter of the B-cell lymphoma-2 (bcl-2) gene contains a highly guaninecytosine-rich region, which has a role in the regulation of bcl-2 transcription. Whereas the guanine-rich region has been the focus of recent studies, little attention has been paid to the cytosine-rich strand. Here we examine the structural transitions of the cytosine-rich sequence by means of acid-base, mole-ratio and melting experiments monitored by molecular absorption, circular dichroism, and NMR spectroscopies. Two intramolecular i-motif structures have been detected in the pH range 2-7, with maximal formation at pH 4 and 6, respectively. At pH 7.6 the majority species has been associated with a hairpin involving Watson-Crick base pairs. Upon addition of the quadruplex-interacting ligand TmPyP4, bcl-2c structures at pH 6.1 and 7.6 yield identical interaction species with stoichiometries 1:2 (DNA:ligand) and logarithms of formation constant 12.4+/-0.2 and 11.7+/-0.1, respectively. The initial i-motif structure at pH 6.1 is lost upon interaction with TmPyP4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs*
  • Base Sequence
  • Circular Dichroism
  • Cytosine / chemistry
  • DNA / chemistry*
  • DNA / genetics
  • Humans
  • Hydrogen-Ion Concentration
  • Ligands
  • Lymphoma, B-Cell / genetics*
  • Lymphoma, B-Cell / pathology
  • Models, Chemical
  • Nuclear Magnetic Resonance, Biomolecular
  • Porphyrins / chemistry
  • Promoter Regions, Genetic*
  • Lösungen
  • Spectrophotometry, Ultraviolet
  • Temperature
  • Thermodynamics

Substances

  • Ligands
  • Porphyrins
  • Lösungen
  • Cytosine
  • DNA