Oxygen metabolism and reactive oxygen species cause chromosomal rearrangements and cell death

doi:10.1073/pnas.0703192104

Oxygen metabolism and reactive oxygen species cause chromosomal rearrangements and cell death

*Centre National de la Recherche Scientifique, Unité Mixte de Recherche 2027, Institut Curie, Bâtiment 110, Centre Universitaire, 91405 Orsay, France; and
^†Ludwig Institute for Cancer Research, Department of Medicine and Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, CA 92093

Contributed by Richard D. Kolodner, April 21, 2007 (received for review February 17, 2007)

Abstract

The absence of Tsa1, a key peroxiredoxin that functions to scavenge H₂O₂ in Saccharomyces cerevisiae, causes the accumulation of a broad spectrum of mutations including gross chromosomal rearrangements (GCRs). Deletion of TSA1 also causes synthetic lethality in combination with mutations in RAD6 and several key genes involved in DNA double-strand break repair. In the present study we investigated the causes of GCRs and cell death in these mutants. tsa1-associated GCRs were independent of the activity of the translesion DNA polymerases ζ, η, and Rev1. Anaerobic growth reduced substantially GCR rates of WT and tsa1 mutants and restored the viability of tsa1 rad6, tsa1 rad51, and tsa1 mre11 double mutants. Anaerobic growth also reduced the GCR rate of rad27, pif1, and rad52 mutants, indicating a role of reactive oxygen species in GCR formation in these mutants. In addition, deletion of TSA1 or H₂O₂ treatment of WT cells resulted in increased formation of Rad52 foci, sites of repair of multiple DNA lesions. H₂O₂ treatment also induced the GCRs. Our results provide in vivo evidence that oxygen metabolism and reactive oxygen species are important sources of DNA damages that can lead to GCRs and lethal effects in S. cerevisiae.

Footnotes

^‡To whom correspondence may be addressed. E-mail: rkolodner{at}ucsd.edu or meng-er.huang{at}curie.u-psud.fr
Author contributions: S.R., R.D.K., and M.-E.H. designed research; S.R., G.F., and M.-E.H. performed research; I.I. and A.M.-H. contributed new reagents/analytic tools; S.R., G.F., R.D.K., and M.-E.H. analyzed data; and R.D.K. and M.-E.H. wrote the paper.
The authors declare no conflict of interest.
Abbreviations:

GCR,

gross chromosomal rearrangement;

ROS,

reactive oxygen species;

DSB,

dsDNA break;

TLS,

translesion DNA synthesis;

Canr,

canavanine-resistant;

Canr-5FOAr,

canavanine- and 5-fluoroorotic acid-resistant;

YPD,

yeast extract/peptone/dextrose;

SC,

synthetic complete.