Generation of double Holliday junction DNAs and their dissolution/resolution within a chromatin context

More about Open Access at the Crick

Abstract

SignificanceWe devised a method involving DNAzyme self-cleavage for the preparation of DNA molecules containing double Holliday junctions (dHJs) that are separated by 746 bp. The DNAs can be prepared in large amounts suitable for in vitro analysis of enzymes required for the processing of recombination intermediates. We show that the dHJ DNA is dissolved efficiently by the BLM-TopIIIα-RMI1-RMI2 (BTRR) complex to produce noncrossover products, whereas nucleolytic resolution by GEN or SMX results in both crossover and noncrossover products. Following nucleosome assembly, the chromatinized dHJ structures are preferentially dissolved by BTRR. These new Holliday junction substrates will provide a valuable resource for the mechanistic analyses of the way in which recombination intermediates are processed.

Journal details

Volume 119
Issue number 18
Pages e2123420119
Available online
Publication date

Crick authors

Crick First author
Crick Corresponding author