A Flp-nick System to Study Repair of a Single Protein-bound Nick in Vivo

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2009 Sep 15

PMID 19749762

Citations 34

Authors

Ida Nielsen

Iben Bach Bentsen

Michael Lisby

Sabine Hansen

Kamilla Mundbjerg

Anni H Andersen

Lotte Bjergbaek

Affiliations

Soon will be listed here.

Abstract

We present the Flp-nick system, which allows introduction of a protein-bound nick at a single genomic site in Saccharomyces cerevisiae and thus mimics a stabilized topoisomerase I-DNA cleavage complex. We took advantage of a mutant Flp recombinase that can introduce a nick at a specific Flp recombinase recognition target site that has been integrated in the yeast genome. The genetic requirement for cells to cope with this insult is the same as for cells treated with camptothecin, which traps topoisomerase I-DNA cleavage complexes genome-wide. Hence, a single protein-bound nick is enough to kill cells if functional repair pathways are lacking. The Flp-nick system can be used to dissect repair, checkpoint and replication fork management pathways activated by a single genomic insult, and it allows the study of events at the damage site, which so far has been impossible to address.

Citing Articles

Two-ended recombination at a Flp-nickase-broken replication fork.

Elango R, Nilavar N, Li A, Nguyen D, Rass E, Duffey E Mol Cell. 2024; 85(1):78-90.e3.

PMID: 39631396 PMC: 11733529. DOI: 10.1016/j.molcel.2024.11.006.

Repair of replication-dependent double-strand breaks differs between the leading and lagging strands.

Kimble M, Sane A, Reid R, Johnson M, Rothstein R, Symington L Mol Cell. 2024; 85(1):61-77.e6.

PMID: 39631395 PMC: 11698654. DOI: 10.1016/j.molcel.2024.10.032.

One-ended and two-ended breaks at nickase-broken replication forks.

Scully R, Walter J, Nussenzweig A DNA Repair (Amst). 2024; 144:103783.

PMID: 39504607 PMC: 11681922. DOI: 10.1016/j.dnarep.2024.103783.

Induction of homologous recombination by site-specific replication stress.

Triplett M, Johnson M, Symington L DNA Repair (Amst). 2024; 142:103753.

PMID: 39190984 PMC: 11425181. DOI: 10.1016/j.dnarep.2024.103753.

PARP1-dependent DNA-protein crosslink repair.

Fabian Z, Kakulidis E, Hendriks I, Kuhbacher U, Larsen N, Oliva-Santiago M Nat Commun. 2024; 15(1):6641.

PMID: 39103378 PMC: 11300803. DOI: 10.1038/s41467-024-50912-x.

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