Gel electrophoretic analysis of the geometry of a DNA four-way junction

Julia Promisel Cooper, Paul J Hagerman

Research output: Contribution to journalArticlepeer-review

157 Scopus citations

Abstract

Branched DNA molecules (Holliday structures) are believed to be key intermediates in the process of homologous genetic recombination. However, despite the importance of such structures, their transient nature makes it difficult to analyze their physical properties. In an effort to evaluate several models for the geometry of such branched molecules, a stable, synthetic DNA four-way junction has been constructed. The geometry of the synthetic junction has been probed by gel electrophoresis, utilizing the fact that bent DNA molecules demonstrate reduced mobilities on polyacrylamide gels to an extent that varies with the degree of the bend angle. From the synthetic four-way junction, we have produced a set of molecules in which all combinations of two junction arms have been extended by 105 base-pairs. The electrophoretic mobilities of the extended junctions differ in a manner which indicates that the junction is not a completely flexible structure; nor is it tetrahedral or planar-tetragonal. Instead, the four strands that comprise the DNA four-way junction are structurally non-equivalent. The significance of these observations with regard to previous models for four-way junction geometry is discussed.

Original languageEnglish (US)
Pages (from-to)711-719
Number of pages9
JournalJournal of Molecular Biology
Volume198
Issue number4
DOIs
StatePublished - Dec 20 1987
Externally publishedYes

ASJC Scopus subject areas

  • Virology

Fingerprint Dive into the research topics of 'Gel electrophoretic analysis of the geometry of a DNA four-way junction'. Together they form a unique fingerprint.

Cite this