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Frequency of DNA end joining in trans is not determined by the predamage spatial proximity of double-strand breaks in yeast.
Proc Natl Acad Sci U S A. 2019 05 07; 116(19):9481-9490.PN

Abstract

DNA double-strand breaks (DSBs) are serious genomic insults that can lead to chromosomal rearrangements if repaired incorrectly. To gain insight into the nuclear mechanisms contributing to these rearrangements, we developed an assay in yeast to measure cis (same site) vs. trans (different site) repair for the majority process of precise nonhomologous end joining (NHEJ). In the assay, the HO endonuclease gene is placed between two HO cut sites such that HO expression is self-terminated upon induction. We further placed an additional cut site in various genomic loci such that NHEJ in trans led to expression of a LEU2 reporter gene. Consistent with prior reports, cis NHEJ was more efficient than trans NHEJ. However, unlike homologous recombination, where spatial distance between a single DSB and donor locus was previously shown to correlate with repair efficiency, trans NHEJ frequency remained essentially constant regardless of the position of the two DSB loci, even when they were on the same chromosome or when two trans repair events were put in competition. Repair of similar DSBs via single-strand annealing of short terminal direct repeats showed substantially higher repair efficiency and trans repair frequency, but still without a strong correlation of trans repair to genomic position. Our results support a model in which yeast cells mobilize, and perhaps compartmentalize, multiple DSBs in a manner that no longer reflects the predamage position of two broken loci.

Authors+Show Affiliations

Department of Pathology, University of Michigan, Ann Arbor, MI 48109.Department of Pathology, University of Michigan, Ann Arbor, MI 48109; wilsonte@umich.edu. Department of Human Genetics, University of Michigan, Ann Arbor, MI 48109.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

31019070

Citation

Sunder, Sham, and Thomas E. Wilson. "Frequency of DNA End Joining in Trans Is Not Determined By the Predamage Spatial Proximity of Double-strand Breaks in Yeast." Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 19, 2019, pp. 9481-9490.
Sunder S, Wilson TE. Frequency of DNA end joining in trans is not determined by the predamage spatial proximity of double-strand breaks in yeast. Proc Natl Acad Sci U S A. 2019;116(19):9481-9490.
Sunder, S., & Wilson, T. E. (2019). Frequency of DNA end joining in trans is not determined by the predamage spatial proximity of double-strand breaks in yeast. Proceedings of the National Academy of Sciences of the United States of America, 116(19), 9481-9490. https://doi.org/10.1073/pnas.1818595116
Sunder S, Wilson TE. Frequency of DNA End Joining in Trans Is Not Determined By the Predamage Spatial Proximity of Double-strand Breaks in Yeast. Proc Natl Acad Sci U S A. 2019 05 7;116(19):9481-9490. PubMed PMID: 31019070.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Frequency of DNA end joining in trans is not determined by the predamage spatial proximity of double-strand breaks in yeast. AU - Sunder,Sham, AU - Wilson,Thomas E, Y1 - 2019/04/24/ PY - 2019/4/26/pubmed PY - 2020/3/17/medline PY - 2019/4/26/entrez KW - genome rearrangement KW - homologous recombination KW - nonhomologous end joining KW - single-strand annealing KW - translocation SP - 9481 EP - 9490 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc Natl Acad Sci U S A VL - 116 IS - 19 N2 - DNA double-strand breaks (DSBs) are serious genomic insults that can lead to chromosomal rearrangements if repaired incorrectly. To gain insight into the nuclear mechanisms contributing to these rearrangements, we developed an assay in yeast to measure cis (same site) vs. trans (different site) repair for the majority process of precise nonhomologous end joining (NHEJ). In the assay, the HO endonuclease gene is placed between two HO cut sites such that HO expression is self-terminated upon induction. We further placed an additional cut site in various genomic loci such that NHEJ in trans led to expression of a LEU2 reporter gene. Consistent with prior reports, cis NHEJ was more efficient than trans NHEJ. However, unlike homologous recombination, where spatial distance between a single DSB and donor locus was previously shown to correlate with repair efficiency, trans NHEJ frequency remained essentially constant regardless of the position of the two DSB loci, even when they were on the same chromosome or when two trans repair events were put in competition. Repair of similar DSBs via single-strand annealing of short terminal direct repeats showed substantially higher repair efficiency and trans repair frequency, but still without a strong correlation of trans repair to genomic position. Our results support a model in which yeast cells mobilize, and perhaps compartmentalize, multiple DSBs in a manner that no longer reflects the predamage position of two broken loci. SN - 1091-6490 UR - https://www.unboundmedicine.com/medline/citation/31019070/Frequency_of_DNA_end_joining_in_trans_is_not_determined_by_the_predamage_spatial_proximity_of_double_strand_breaks_in_yeast_ L2 - http://www.pnas.org/cgi/pmidlookup?view=long&pmid=31019070 DB - PRIME DP - Unbound Medicine ER -