title: Determination of gross chromosomal rearrangement rates.
abstract: Cells devote a significant amount of metabolism to maintaining the stability of their genome and to preventing inappropriate chromosomal rearrangements that are characteristic of many cancers. A simple genetic assay using haploid derivatives of the yeast Saccharomyces cerevisiae provides a means to quantitatively measure the rate at which gross chromosomal rearrangements (GCRs) accumulate in different genetic backgrounds. This assay measures the rate of simultaneous inactivation of CAN1 and URA3 markers placed on a nonessential end of a yeast chromosome and in principle can be implemented in any haploid strain. Rearrangements detected with this assay include broken chromosomes healed by de novo telomere additions and a spectrum of inter- and intrachromosomal fusion events. The GCR assay allows for detailed analysis of the contributions of individual genes and different pathways in the suppression of genomic instability.
authors.affiliation: Ludwig Institute for Cancer Research, La Jolla, CA 92093-2385, USA. cdputnam@ucsd.edu
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Putnam
authors.forename: Richard D
authors.initials: RD
authors.lastname: Kolodner
mesh_terms: D026923:Amino Acid Transport Systems, Basic / Q000235:genetics; D043171:Chromosomal Instability; D015825:Chromosomes, Fungal; D004271:DNA, Fungal / Q000235:genetics*; D015321:Gene Rearrangement; D020868:Gene Silencing; D005821:Genetic Techniques; D042822:Genomic Instability; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D026923:Amino Acid Transport Systems, Basic; C502212:CAN1 protein, S cerevisiae; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; C554167:URA3 protein, S cerevisiae
references.citation: Mol Cell Biol. 2006 Jul;26(14):5406-20
references.pmid: 16809776
references.citation: Nat Genet. 1999 Sep;23(1):81-5
references.pmid: 10471504
references.citation: Methods. 2007 Feb;41(2):168-76
references.pmid: 17189859
references.citation: J Genet. 1949 Dec;49(3):264-85
references.pmid: 24536673
references.citation: Science. 2002 Jul 26;297(5581):552-7
references.pmid: 12142524
references.citation: Cell. 2001 Aug 10;106(3):275-86
references.pmid: 11509177
references.citation: Nature. 2009 Aug 20;460(7258):984-9
references.pmid: 19641493
references.citation: PLoS One. 2009 Jul 28;4(7):e6389
references.pmid: 19636429
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):9039-44
references.pmid: 15184655
references.citation: Cell. 2001 Feb 9;104(3):397-408
references.pmid: 11239397
references.citation: Methods Enzymol. 2006;409:462-76
references.pmid: 16793418
references.citation: Nat Genet. 2001 Jan;27(1):113-6
references.pmid: 11138010
references.citation: Nat Genet. 2005 Jul;37(7):750-5
references.pmid: 15965476