PMID- 32840803
OWN - NLM
STAT- MEDLINE
DCOM- 20210316
LR  - 20220102
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 2153
DP  - 2021
TI  - Physical and Genetic Assays for the Study of DNA Joint Molecules Metabolism and 
      Multi-invasion-Induced Rearrangements in S. cerevisiae.
PG  - 535-554
LID - 10.1007/978-1-0716-0644-5_36 [doi]
AB  - DNA double-strand breaks (DSBs) are genotoxic lesions that can be repaired in a 
      templated fashion by homologous recombination (HR). HR is a complex pathway that 
      involves the formation of DNA joint molecules (JMs) containing heteroduplex DNA. 
      Various types of JMs are formed throughout the pathway, including displacement 
      loops (D-loops), multi-invasions (MI), and double Holliday junction 
      intermediates. Dysregulation of JM metabolism in various mutant contexts revealed 
      the propensity of HR to generate repeat-mediated chromosomal rearrangements. 
      Specifically, we recently identified MI-induced rearrangements (MIR), a 
      tripartite recombination mechanism initiated by one end of a DSB that exploits 
      repeated regions to generate rearrangements between intact chromosomal regions. 
      MIR occurs upon MI-JM processing by endonucleases and is suppressed by JM 
      disruption activities. Here, we detail two assays: a physical assay for JM 
      detection in Saccharomyces cerevisiae cells and genetic assays to determine the 
      frequency of MIR in various chromosomal contexts. These assays enable studying 
      the regulation of the HR pathway and the consequences of their defects for 
      genomic instability by MIR.
FAU - Piazza, Aurele
AU  - Piazza A
AD  - Spatial Regulation of Genomes, Institut Pasteur, UMR3525 CNRS, Paris, France.
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, CA, USA.
AD  - Univ Lyon, ENS, UCBL, CNRS, INSERM, Laboratory of Biology and Modelling of the 
      Cell, UMR5239, Lyon, France.
FAU - Rajput, Pallavi
AU  - Rajput P
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, CA, USA.
FAU - Heyer, Wolf-Dietrich
AU  - Heyer WD
AD  - Department of Microbiology and Molecular Genetics, University of California, 
      Davis, CA, USA. wdheyer@ucdavis.edu.
AD  - Department of Molecular and Cellular Biology, University of California, Davis, 
      CA, USA. wdheyer@ucdavis.edu.
LA  - eng
GR  - R01 CA092276/CA/NCI NIH HHS/United States
GR  - R01 GM058015/GM/NIGMS NIH HHS/United States
GR  - R01 GM137751/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (DNA, Fungal)
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Nucleic Acid Heteroduplexes)
RN  - EC 3.6.1 (Holliday junction DNA helicase, E coli)
RN  - EC 3.6.4.- (DNA Helicases)
SB  - IM
MH  - DNA Breaks, Double-Stranded
MH  - DNA Helicases/metabolism
MH  - DNA, Fungal/chemistry/*genetics
MH  - Escherichia coli Proteins/metabolism
MH  - Mutation
MH  - Nucleic Acid Heteroduplexes
MH  - *Recombinational DNA Repair
MH  - Saccharomyces cerevisiae/*genetics/metabolism
PMC - PMC8115024
MID - NIHMS1682192
OTO - NOTNLM
OT  - D-loop
OT  - DNA repair
OT  - Genomic instability
OT  - Homologous recombination
OT  - MIR
OT  - Multi-invasions
EDAT- 2020/08/26 06:00
MHDA- 2021/03/17 06:00
PMCR- 2022/01/01
CRDT- 2020/08/26 06:00
PHST- 2020/08/26 06:00 [entrez]
PHST- 2020/08/26 06:00 [pubmed]
PHST- 2021/03/17 06:00 [medline]
PHST- 2022/01/01 00:00 [pmc-release]
AID - 10.1007/978-1-0716-0644-5_36 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2021;2153:535-554. doi: 10.1007/978-1-0716-0644-5_36.