Determination of gross chromosomal rearrangement rates.
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.
Putnam
Christopher D
CD
Ludwig Institute for Cancer Research, La Jolla, CA 92093-2385, USA. cdputnam@ucsd.edu
Kolodner
Richard D
RD
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
D026923:Amino Acid Transport Systems, Basic; C502212:CAN1 protein, S cerevisiae; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; C554167:URA3 protein, S cerevisiae
Mol Cell Biol. 2006 Jul;26(14):5406-20
16809776
Nat Genet. 1999 Sep;23(1):81-5
10471504
Methods. 2007 Feb;41(2):168-76
17189859
J Genet. 1949 Dec;49(3):264-85
24536673
Science. 2002 Jul 26;297(5581):552-7
12142524
Cell. 2001 Aug 10;106(3):275-86
11509177
Nature. 2009 Aug 20;460(7258):984-9
19641493
PLoS One. 2009 Jul 28;4(7):e6389
19636429
Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):9039-44
15184655
Cell. 2001 Feb 9;104(3):397-408
11239397
Methods Enzymol. 2006;409:462-76
16793418
Nat Genet. 2001 Jan;27(1):113-6
11138010
Nat Genet. 2005 Jul;37(7):750-5
15965476