Tags

Type your tag names separated by a space and hit enter

Two different types of double-strand breaks in Saccharomyces cerevisiae are repaired by similar RAD52-independent, nonhomologous recombination events.
Mol Cell Biol. 1994 Feb; 14(2):1293-301.MC

Abstract

In haploid rad52 Saccharomyces cerevisiae strains unable to undergo homologous recombination, a chromosomal double-strand break (DSB) can be repaired by imprecise rejoining of the broken chromosome ends. We have used two different strategies to generate broken chromosomes: (i) a site-specific DSB generated at the MAT locus by HO endonuclease cutting or (ii) a random DSB generated by mechanical rupture during mitotic segregation of a conditionally dicentric chromosome. Broken chromosomes were repaired by deletions that were highly variable in size, all of which removed more sequences than was required either to prevent subsequent HO cleavage or to eliminate a functional centromere, respectively. The junction of the deletions frequently occurred where complementary strands from the flanking DNA could anneal to form 1 to 5 bp, although 12% (4 of 34) of the events appear to have occurred by blunt-end ligation. These types of deletions are very similar to the junctions observed in the repair of DSBs by mammalian cells (D. B. Roth and J. H. Wilson, Mol. Cell. Biol. 6:4295-4304, 1986). When a high level of HO endonuclease, expressed in all phases of the cell cycle, was used to create DSBs, we also recovered a large class of very small (2- or 3-bp) insertions in the HO cleavage site. These insertions appear to represent still another mechanism of DSB repair, apparently by annealing and filling in the overhanging 3' ends of the cleavage site. These types of events have also been well documented for vertebrate cells.

Authors+Show Affiliations

Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02254-9110.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8289808

Citation

Kramer, K M., et al. "Two Different Types of Double-strand Breaks in Saccharomyces Cerevisiae Are Repaired By Similar RAD52-independent, Nonhomologous Recombination Events." Molecular and Cellular Biology, vol. 14, no. 2, 1994, pp. 1293-301.
Kramer KM, Brock JA, Bloom K, et al. Two different types of double-strand breaks in Saccharomyces cerevisiae are repaired by similar RAD52-independent, nonhomologous recombination events. Mol Cell Biol. 1994;14(2):1293-301.
Kramer, K. M., Brock, J. A., Bloom, K., Moore, J. K., & Haber, J. E. (1994). Two different types of double-strand breaks in Saccharomyces cerevisiae are repaired by similar RAD52-independent, nonhomologous recombination events. Molecular and Cellular Biology, 14(2), 1293-301.
Kramer KM, et al. Two Different Types of Double-strand Breaks in Saccharomyces Cerevisiae Are Repaired By Similar RAD52-independent, Nonhomologous Recombination Events. Mol Cell Biol. 1994;14(2):1293-301. PubMed PMID: 8289808.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Two different types of double-strand breaks in Saccharomyces cerevisiae are repaired by similar RAD52-independent, nonhomologous recombination events. AU - Kramer,K M, AU - Brock,J A, AU - Bloom,K, AU - Moore,J K, AU - Haber,J E, PY - 1994/2/1/pubmed PY - 1994/2/1/medline PY - 1994/2/1/entrez SP - 1293 EP - 301 JF - Molecular and cellular biology JO - Mol Cell Biol VL - 14 IS - 2 N2 - In haploid rad52 Saccharomyces cerevisiae strains unable to undergo homologous recombination, a chromosomal double-strand break (DSB) can be repaired by imprecise rejoining of the broken chromosome ends. We have used two different strategies to generate broken chromosomes: (i) a site-specific DSB generated at the MAT locus by HO endonuclease cutting or (ii) a random DSB generated by mechanical rupture during mitotic segregation of a conditionally dicentric chromosome. Broken chromosomes were repaired by deletions that were highly variable in size, all of which removed more sequences than was required either to prevent subsequent HO cleavage or to eliminate a functional centromere, respectively. The junction of the deletions frequently occurred where complementary strands from the flanking DNA could anneal to form 1 to 5 bp, although 12% (4 of 34) of the events appear to have occurred by blunt-end ligation. These types of deletions are very similar to the junctions observed in the repair of DSBs by mammalian cells (D. B. Roth and J. H. Wilson, Mol. Cell. Biol. 6:4295-4304, 1986). When a high level of HO endonuclease, expressed in all phases of the cell cycle, was used to create DSBs, we also recovered a large class of very small (2- or 3-bp) insertions in the HO cleavage site. These insertions appear to represent still another mechanism of DSB repair, apparently by annealing and filling in the overhanging 3' ends of the cleavage site. These types of events have also been well documented for vertebrate cells. SN - 0270-7306 UR - https://www.unboundmedicine.com/medline/citation/8289808/Two_different_types_of_double_strand_breaks_in_Saccharomyces_cerevisiae_are_repaired_by_similar_RAD52_independent_nonhomologous_recombination_events_ L2 - https://journals.asm.org/doi/10.1128/mcb.14.2.1293-1301.1994?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -