Spontaneous DNA Damage in Saccharomyces cerevisiae Elicits Phenotypic Properties Similar to Cancer Cells

Evert, Barbara A.; Salmon, Tiffany B.; Song, Binwei; Liu, Jingjing; Siede, Wolfram; Doetsch, Paul W.

doi:10.1074/jbc.m400468200

Cited by 60 publications

(65 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To investigate the role of oxidative DNA damage in the induction of genetic instability, we constructed a series of strains with compromised (14). Thus, biologically relevant, elevated levels of endogenously produced oxidative DNA damage can be achieved when several BER genes are disrupted simultaneously (16). To exclude the possibility of selection for additional mutations and modifications as a consequence of multiple gene deletions in a haploid background, we consecutively eliminated the excision repair genes and the TSA1 gene in a wild-type diploid strain containing the reporter systems for measuring genetic/genomic instability (see Materials and Methods).…”

Section: Resultsmentioning

confidence: 99%

“…A BER-and NER-defective strain (an ntg1⌬ ntg2⌬ apn1⌬ rad1⌬ quadruple mutant) provides a unique tool for delineating the mechanisms of cellular responses to such stress, because the levels of ROS and chronic oxidative DNA damage in this strain are remarkably high, approximately equivalent to a 50% lethal acute-exposure dose (3 mM) of hydrogen peroxide (16). Such DNA repair-deficient cells, harboring high levels of oxidative DNA damage and elevated levels of ROS, display high mutation and recombination frequencies, as well as other abnormalities, including slow growth and extreme sensitivity to DNA-damaging agents (16,47). These findings indicate that if the levels of DNA damage exceed the capacity of the major excision repair pathways (BER and NER) to maintain the integrity of the genome, damage tolerance pathway-mediated events confer a state of genetic instability.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Chronic Oxidative DNA Damage Due to DNA Repair Defects Causes Chromosomal Instability in Saccharomyces cerevisiae

Degtyareva

Chen²,

Mieczkowski

et al. 2008

Molecular and Cellular Biology

Self Cite

View full text Add to dashboard Cite

Oxidative DNA damage is likely to be involved in the etiology of cancer and is thought to accelerate tumorigenesis via increased mutation rates. However, the majority of malignant cells acquire a specific type of genomic instability characterized by large-scale genomic rearrangements, referred to as chromosomal instability (CIN). The molecular mechanisms underlying CIN are not entirely understood. We utilized Saccharomyces cerevisiae as a model system to delineate the relationship between genotoxic stress and CIN. It was found that elevated levels of chronic, unrepaired oxidative DNA damage caused chromosomal aberrations at remarkably high frequencies under both selective and nonselective growth conditions. In this system, exceeding the cellular capacity to appropriately manage oxidative DNA damage resulted in a "gain-of-CIN" phenotype and led to profound karyotypic instability. These results illustrate a novel mechanism for genome destabilization that is likely to be relevant to human carcinogenesis.Genetic instability is a common feature acquired by cancer cells, enabling the development and progression of tumors (23). Events resulting in chromosomal instability (CIN), such as amplifications and deletions of large segments of DNA, reciprocal and nonreciprocal translocations, aneuploidy, and polyploidy, constitute the large-scale genomic aberrations that characterize the majority of human cancer cells and are thought to accelerate carcinogenesis (37). The molecular mechanisms underlying CIN remain to be elucidated and are of profound importance for understanding tumorigenesis (45). Significant effort has been invested in the search for genes that when inactivated result in genome destabilization and subsequent, rapid tumor development. Such genes include those controlling the mitotic checkpoint and sister-chromatid separation (26). Most of the studies investigating the mechanisms, timing, and relevance of CIN in tumorigenesis have been focused on genetic or epigenetic aspects of CIN. Little is known about environmental factors that may cause and/or promote CIN during tumor development.Exogenous (environmental) and endogenous (intracellular) oxidative DNA damage is considered to play an important role in cancer etiology (25). Chronic inflammation involving the release of free radicals by leukocytes, acquired through chemical insults or viral and bacterial infections, is thought to contribute to about one in four cancers worldwide (10). Strong evidence for a direct and specific role of reactive oxygen species (ROS) in the oncogenic transformation of cells has been provided recently by the finding that the activation of two major oncogenes, the Ras and Myc oncogenes, increases intracellular levels of ROS and induces DNA damage and genomic instability (30, 54). Cells have evolved several mechanisms for the prevention and repair of oxidative damage. The base excision repair (BER) pathway is responsible for the removal of a large proportion of oxidative DNA damage, although when this pathway is inactivated or its cap...

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Chronic Oxidative DNA Damage Due to DNA Repair Defects Causes Chromosomal Instability in Saccharomyces cerevisiae

Degtyareva

Chen²,

Mieczkowski

et al. 2008

Molecular and Cellular Biology

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, a genome-wide screen in S. cerevisiae for genes that suppress the accumulation of mutations identified oxidized guanine and uracil in DNA as major contributors to spontaneous mutagenesis (64), and if unrepaired, these endogenous DNA base lesions clearly contribute to neoplastic transformation of mammalian cells (4, 70,97,106,127,134,154,157). When actual levels of unrepaired spontaneous DNA base damage are carefully evaluated for their biological consequences, in compound mutant strains, even yeast exhibit phenotypic properties similar to those of cancer cells (39). So the base-excision repair field has in many ways come full circle and we might think again of BER in the context of tumorigenesis and potential cancer therapeutic targets (88), but now from our knowledge of mammalian rather than microbial systems.…”

Section: Resultsmentioning

confidence: 99%

Repair and Genetic Consequences of Endogenous DNA Base Damage in Mammalian Cells

2004

View full text Add to dashboard Cite

Key Words DNA repair, mutagenesis, knock-out mice, uracil in DNA, DNA oxidation ■ Abstract Living organisms dependent on water and oxygen for their existence face the major challenge of faithfully maintaining their genetic material under a constant attack from spontaneous hydrolysis and active oxygen species and from other intracellular metabolites that can modify DNA bases. Repair of endogenous DNA base damage by the ubiquitous base-excision repair pathway largely accounts for the significant turnover of DNA even in nonreplicating cells, and must be sufficiently accurate and efficient to preserve genome stability compatible with long-term cellular viability.

show abstract

“…To investigate the relationship between oxidative stress and oxidative mtDNA damage, we utilized a mitochondrial gene-specific DNA damage detection assay that measures the amount of Ntg1p-recognizable lesions in a 4.4-kb fragment of the mitochondrial genome. Total cellular DNA was obtained using a method that minimizes introduction of adventitious oxidative DNA damage (15). DNA was subsequently treated with Ntg1p, a BER DNA glycosylase with associated AP lyase activity that primarily recognizes and cleaves DNA containing oxidative pyrimidine base damage and abasic sites (56).…”

Section: Resultsmentioning

confidence: 99%

“…This electrophoretic assay determines the levels of Ntg1p recognizable lesions present within a coding region of the mitochondrial genome and is an adaptation of a similar method previously used by our group for measuring oxidative nuclear DNA damage (15). Tenmicrogram aliquots of genomic DNA were digested with NdeI for 4 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Oxidative DNA Damage Causes Mitochondrial Genomic Instability in Saccharomyces cerevisiae

Doudican¹,

Song²,

Shadel

et al. 2005

Molecular and Cellular Biology

Self Cite

133

110

View full text Add to dashboard Cite

Mitochondria contain their own genome, the integrity of which is required for normal cellular energy metabolism. Reactive oxygen species (ROS) produced by normal mitochondrial respiration can damage cellular macromolecules, including mitochondrial DNA (mtDNA), and have been implicated in degenerative diseases, cancer, and aging. We developed strategies to elevate mitochondrial oxidative stress by exposure to antimycin and H 2 O 2 or utilizing mutants lacking mitochondrial superoxide dismutase (sod2⌬). Experiments were conducted with strains compromised in mitochondrial base excision repair (ntg1⌬) and oxidative damage resistance (pif1⌬) in order to delineate the relationship between these pathways. We observed enhanced ROS production, resulting in a direct increase in oxidative mtDNA damage and mutagenesis. Repair-deficient mutants exposed to oxidative stress conditions exhibited profound genomic instability. Elimination of Ntg1p and Pif1p resulted in a synergistic corruption of respiratory competency upon exposure to antimycin and H 2 O 2 . Mitochondrial genomic integrity was substantially compromised in ntg1⌬ pif1⌬ sod2⌬ strains, since these cells exhibit a total loss of mtDNA. A stable respiration-defective strain, possessing a normal complement of mtDNA damage resistance pathways, exhibited a complete loss of mtDNA upon exposure to antimycin and H 2 O 2 . This loss was preventable by Sod2p overexpression. These results provide direct evidence that oxidative mtDNA damage can be a major contributor to mitochondrial genomic instability and demonstrate cooperation of Ntg1p and Pif1p to resist the introduction of lesions into the mitochondrial genome.

show abstract

Spontaneous DNA Damage in Saccharomyces cerevisiae Elicits Phenotypic Properties Similar to Cancer Cells

Abstract: To determine the spectrum of effects elicited by specific levels of spontaneous DNA damage, a series of isogenic Saccharomyces cerevisiae strains defective in base excision repair (BER) and nucleotide excision repair (NER) were analyzed. In log phase of growth, when com-

Cited by 60 publications

References 44 publications

Chronic Oxidative DNA Damage Due to DNA Repair Defects Causes Chromosomal Instability in Saccharomyces cerevisiae

Chronic Oxidative DNA Damage Due to DNA Repair Defects Causes Chromosomal Instability in Saccharomyces cerevisiae

Repair and Genetic Consequences of Endogenous DNA Base Damage in Mammalian Cells

Oxidative DNA Damage Causes Mitochondrial Genomic Instability in Saccharomyces cerevisiae

Contact Info

Product

Resources

About