Interaction of linear homologous DNA duplexes via Holliday junction formation

Interaction of linear homologous DNA duplexes via Holliday junction formation

Interaction of linear homologous DNA duplexes by formation of Holliday junctions was revealed by electrophoresis and confirmed by electron microscopy. The phenomenon was demonstrated using a model of five purified PCR products of different size and sequence. The double‐stranded structure of interact...

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Bibliographic Details
Published in:European journal of biochemistry Vol. 268; no. 1; pp. 7 - 14
Main Authors: Yakubovskaya, Marianna G., Neschastnova, Anna A., Humphrey, Karen E., Babon, Jeff J., Popenko, Vladimir I., Smith, Margaret J., Lambrinakos, Andrianna, Lipatova, Zhanna V., Dobrovolskaia, Marina A., Cappai, Roberto, Masters, Colin L., Belitsky, Gennady A., Cotton, Richard G.
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Science Ltd 01-01-2001
Subjects:
Animals
branch migration
DNA - chemistry
DNA - metabolism
DNA association
DNA complex
duplex interaction
Holliday junction
Humans
Nucleic Acid Conformation
Polymerase Chain Reaction
Sequence Homology, Nucleic Acid
Tumor Suppressor Protein p53 - genetics
Tumor Suppressor Protein p53 - metabolism
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Summary:Interaction of linear homologous DNA duplexes by formation of Holliday junctions was revealed by electrophoresis and confirmed by electron microscopy. The phenomenon was demonstrated using a model of five purified PCR products of different size and sequence. The double‐stranded structure of interacting DNA fragments was confirmed using several consecutive purifications, S1‐nuclease analysis, and electron microscopy. Formation of Holliday junctions depends on DNA concentration. A thermodynamic equilibrium between duplexes and Holliday junctions was shown. We propose that homologous duplex interaction is initiated by nucleation of several dissociated terminal base pairs of two fragments. This process is followed by branch migration creating a population of Holliday junctions with the branch point at different sites. Finally, Holliday junctions are resolved via branch migration to new or previously existing duplexes. The phenomenon is a new property of DNA. This type of DNA–DNA interaction may contribute to the process of Holliday junction formation in vivo controlled by DNA conformation and DNA–protein interactions. It is of practical significance for optimization of different PCR‐based methods of gene analysis, especially those involving heteroduplex formation.
ISSN:0014-2956
1432-1033
DOI:10.1046/j.1432-1327.2001.01861.x