Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 426, Issue 24, Pages 3946-3959Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2014.10.008
Keywords
DNA recombination and repair; Holliday junction resolution; Chaetomium thermophilum; FEN1; thermophilic proteins
Categories
Funding
- Cancer Research UK [C28/A4959]
- Wellcome Trust [0770121Z/05/Z]
- BBSRC [BB/E001777/1] Funding Source: UKRI
- EPSRC [EP/J017094/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E001777/1] Funding Source: researchfish
- Cancer Research UK [11722] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/J017094/1] Funding Source: researchfish
Ask authors/readers for more resources
Processing of Holliday junctions is essential in recombination. We have identified the gene for the junction-resolving enzyme GEN1 from the thermophilic fungus Chaetomium thermophilum and expressed the N-terminal 487-amino-acid section. The protein is a nuclease that is highly selective for four-way DNA junctions, cleaving 1 nt 3' to the point of strand exchange on two strands symmetrically disposed about a diagonal axis. CtGEN1 binds to DNA junctions as a discrete homodimer with nanomolar affinity. Analysis of the kinetics of cruciform cleavage shows that cleavage of the second strand occurs an order of magnitude faster than the first cleavage so as to generate a productive resolution event. All these properties are closely similar to those described for bacterial, phage and mitochondrial junction-resolving enzymes. CtGEN1 is also similar in properties to the human enzyme but lacks the problems with aggregation that currently prevent detailed analysis of the latter protein. CtGEN1 is thus an excellent enzyme with which to engage in biophysical and structural analysis of eukaryotic GEN1. (C) 2014 MRC Laboratory of Molecular Biology. Published by Elsevier Ltd.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available