Biochimie. 2009 Jul;91(7):894-902. Epub 2009 May 3.

Targeting the G-quadruplex-forming region near the P1 promoter in the human BCL-2 gene with the cationic porphyrin TMPyP4 and with the complementary C-rich strand.

Del Toro M, Bucek P, Aviñó A, Jaumot J, González C, Eritja R, Gargallo R.

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Department of Analytical Chemistry, University of Barcelona, Marti i Franques 1-11, E-08028 Barcelona, Spain.

Abstract

The B-cell lymphoma-2 (bcl-2) gene contains a region that has been implicated in the regulation of bcl-2 gene expression. This region can form G-quadruplex structures in solution [J.X. Dai, T.S. Dexheimer, D. Chen, M. Carver, A. Ambrus, R.A. Jones, D.Z. Yang, An intramolecular G-quadruplex structure with mixed parallel/antiparallel G-strands formed in the human BCL-2 promoter region in solution, J. Am. Chem. Soc. 128 (2006) 1096-1098.]. Here, we examined the acid-base and conformational equilibria of this G-quadruplex-forming region (BCL2G), as well as its interaction with both the porphyrin TMPyP4 and with the complementary C-rich strand. We used molecular absorption and circular dichroism techniques, in tandem with multivariate analysis tools. The results revealed the formation of an interaction complex BCL2G:TMPyP4 with a stoichiometry of 1:2 and an equilibrium constant equal to 5.0 (+/-2.3) x 10(13) M(-2). Addition of the complementary C-rich strand to BCL2G induces the predominant formation of the Watson-Crick double-helix with an equilibrium constant equal to 10(7.7) M(-1) (at pH 7.1). Finally, the pH-induced formation of quadruplex structures from the Watson-Crick double-helix is characterized.

PMID:: 19401211; [PubMed - indexed for MEDLINE]

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