Optimization of alternate-strand triple helix formation at the 5'CpG3' and 5'GpC3' junction steps.

Abstract:

:Oligonucleotide-directed triple helix formation normally requires a long tract of oligopyrimidine.oligopurine sequence. This limitation can be partially overcome by alternate-strand triple helix (or switch triple helix) formation which enables recognition of alternating oligopurine/oligopyrimidine sequences. The present work is devoted to the optimization of switch triple helix formation at the 5'CpG3' and 5'GpC3' junction steps by combination of base triplets in Hoogsteen and in reverse Hoogsteen configurations. Rational design by molecular mechanics was first carried out to study the geometrical constraints at different junction steps and to propose a "switch code" which would optimize the interactions at junctions. These predictions were further checked and validated experimentally by gel retardation and DNase I footprinting assays. It was shown that the choice of an appropriate linker nucleotide in the switching third strand plays an important role in the interaction between oligonucleotides and alternating oligopurine/oligopyrimidine target sequences at different junctions: (i) the addition of a cytosine at the junction level in the oligonucleotide optimizes the crossover at the 5'CpG3' junction, whereas (ii) the best crossover at the 5'GpC3' junction step is achieved without any additional nucleotide. These results provide a useful guideline to extend double-stranded DNA sequence recognition by switch triple helix formation.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Marchand C,Sun JS,Bailly C,Waring MJ,Garestier T,Hélène C

doi

10.1021/bi980618+

subject

Has Abstract

pub_date

1998-09-22 00:00:00

pages

13322-9

issue

38

eissn

0006-2960

issn

1520-4995

pii

bi980618+

journal_volume

37

pub_type

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