Extensive DNA end processing by exo1 and sgs1 inhibits break-induced replication.

PLoS Genetics
Vanessa A Marrero, L S Symington

Abstract

Homology-dependent repair of DNA double-strand breaks (DSBs) by gene conversion involves short tracts of DNA synthesis and limited loss of heterozygosity (LOH). For DSBs that present only one end, repair occurs by invasion into a homologous sequence followed by replication to the end of the chromosome resulting in extensive LOH, a process called break-induced replication (BIR). We developed a BIR assay in Saccharomyces cerevisiae consisting of a plasmid with a telomere seeding sequence separated from sequence homologous to chromosome III by an I-SceI endonuclease recognition site. Following cleavage of the plasmid by I-SceI in vivo, de novo telomere synthesis occurs at one end of the vector, and the other end invades at the homologous sequence on chromosome III and initiates replication to the end of the chromosome to generate a stable chromosome fragment (CF). BIR was infrequent in wild-type cells due to degradation of the linearized vector. However, in the exo1Delta sgs1Delta mutant, which is defective in the 5'-3' resection of DSBs, the frequency of BIR was increased by 39-fold. Extension of the invading end of the plasmid was detected by physical analysis two hours after induction of the I-SceI endonuclease in the wild-type...Continue Reading

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Citations

Jul 7, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jeannine R LaRocqueMaria Jasin
Sep 26, 2014·Nature Communications·Julien HardyMarie-Noëlle Simon
Sep 14, 2011·Annual Review of Genetics·Lorraine S Symington, Jean Gautier
Oct 12, 2010·Nucleic Acids Research·Karim BahmedJohn L Nitiss
Oct 27, 2011·Seminars in Cell & Developmental Biology·Torben R Kasparek, Timothy C Humphrey
Sep 8, 2012·Annals of the New York Academy of Sciences·Anne M CasperKaveri D Rajula
Jun 19, 2013·Cell Reports·Alessia BalestriniJohn H J Petrini
Aug 9, 2016·Progress in Biophysics and Molecular Biology·Logan R Myler, Ilya J Finkelstein
Jun 15, 2013·Biomolecules·Cynthia J SakofskyAnna Malkova
Feb 20, 2020·Cell & Bioscience·Jia YuZhenkun Lou
Jun 25, 2015·Microbiology Spectrum·Cheng-Sheng Lee, James E Haber
Aug 20, 2014·Biochemistry·Timothy J Herdendorf, Scott W Nelson
May 26, 2016·Genes & Development·Priyanka Verma, Roger A Greenberg

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Methods Mentioned

BETA
PCR
electrophoresis

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