scholarly journals Shared and distinct roles of Esc2 and Mms21 in suppressing genome rearrangements and regulating intracellular sumoylation

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247132
Author(s):  
Raymond T. Suhandynata ◽  
Yong-Qi Gao ◽  
Ann L. Zhou ◽  
Yusheng Yang ◽  
Pang-Che Wang ◽  
...  
Keyword(s):  
Dna Damage ◽  
Chromosomal Rearrangements ◽  
Dna Helicase ◽  
Growth Defect ◽  
Catalytic Core ◽  
E3 Ligases ◽  
Nucleolar Proteins ◽  
Gross Chromosomal Rearrangements ◽  
Mcm Complex ◽  
Sumo E3 Ligase

Protein sumoylation, especially when catalyzed by the Mms21 SUMO E3 ligase, plays a major role in suppressing duplication-mediated gross chromosomal rearrangements (dGCRs). How Mms21 targets its substrates in the cell is insufficiently understood. Here, we demonstrate that Esc2, a protein with SUMO-like domains (SLDs), recruits the Ubc9 SUMO conjugating enzyme to specifically facilitate Mms21-dependent sumoylation and suppress dGCRs. The D430R mutation in Esc2 impairs its binding to Ubc9 and causes a synergistic growth defect and accumulation of dGCRs with mutations that delete the Siz1 and Siz2 E3 ligases. By contrast, esc2-D430R does not appreciably affect sensitivity to DNA damage or the dGCRs caused by the catalytically inactive mms21-CH. Moreover, proteome-wide analysis of intracellular sumoylation demonstrates that esc2-D430R specifically down-regulates sumoylation levels of Mms21-preferred targets, including the nucleolar proteins, components of the SMC complexes and the MCM complex that acts as the catalytic core of the replicative DNA helicase. These effects closely resemble those caused by mms21-CH, and are relatively unaffected by deleting Siz1 and Siz2. Thus, by recruiting Ubc9, Esc2 facilitates Mms21-dependent sumoylation to suppress the accumulation of dGCRs independent of Siz1 and Siz2.

scholarly journals SUMO E3 ligase Mms21 prevents spontaneous DNA damage induced genome rearrangements

2017 ◽  
Author(s):  
Jason Liang ◽  
Bin-zhong Li ◽  
Alexander P. Tan ◽  
Richard D. Kolodner ◽  
Christopher D. Putnam ◽  
...  
Keyword(s):  
Dna Damage ◽  
E3 Ligase ◽  
Genome Rearrangements ◽  
Dna Helicase ◽  
Dna Lesions ◽  
Genome Maintenance ◽  
New Paradigm ◽  
A Subunit ◽  
Sumo E3 Ligase ◽  
Break Induced Replication

AbstractMms21, a subunit of the Smc5/6 complex, possesses an E3 ligase activity for the Small Ubiquitin-like MOdifier (SUMO), which has a major, but poorly understood role in genome maintenance. Here we show mutations that inactivate the E3 ligase activity of Mms21 cause Rad52- and Pol32-dependent break-induced replication (BIR), which specifically requires the Rrm3 DNA helicase. Interestingly, mutations affecting both Mms21 and the Sgs1 helicase, but not sumoylation of Sgs1, cause further accumulation of genome rearrangements, indicating the distinct roles of Mms21 and Sgs1 in suppressing genome rearrangements. Whole genome sequencing further revealed that the Mre11 endonuclease prevents microhomology-mediated translocations and hairpin-mediated inverted duplications in the mms21 mutant. Consistent with the accumulation of endogenous DNA lesions, mms21 cells accumulate spontaneous Ddc2 foci and display a hyper-activated DNA damage checkpoint. Together, these findings support a new paradigm that Mms21 prevents the accumulation of spontaneous DNA lesions that cause diverse genome rearrangements.

scholarly journals Functions of Saccharomyces cerevisiae 14-3-3 Proteins in Response to DNA Damage and to DNA Replication Stress

Genetics ◽  
2003 ◽  
Vol 165 (4) ◽  
pp. 1717-1732
Author(s):  
Francisca Lottersberger ◽  
Fabio Rubert ◽  
Veronica Baldo ◽  
Giovanna Lucchini ◽  
Maria Pia Longhese
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Damage ◽  
Dna Replication ◽  
Genome Instability ◽  
Chromosomal Rearrangements ◽  
Replication Stress ◽  
Checkpoint Kinases ◽  
Gross Chromosomal Rearrangements ◽  
Dna Damage Checkpoints ◽  
Dna Replication Stress

Abstract Two members of the 14-3-3 protein family, involved in key biological processes in different eukaryotes, are encoded by the functionally redundant Saccharomyces cerevisiae BMH1 and BMH2 genes. We produced and characterized 12 independent bmh1 mutant alleles, whose presence in the cell as the sole 14-3-3 source causes hypersensitivity to genotoxic agents, indicating that Bmh proteins are required for proper response to DNA damage. In particular, the bmh1-103 and bmh1-266 mutant alleles cause defects in G1/S and G2/M DNA damage checkpoints, whereas only the G2/M checkpoint is altered by the bmh1-169 and bmh1-221 alleles. Impaired checkpoint responses correlate with the inability to maintain phosphorylated forms of Rad53 and/or Chk1, suggesting that Bmh proteins might regulate phosphorylation/dephosphorylation of these checkpoint kinases. Moreover, several bmh1 bmh2Δ mutants are defective in resuming DNA replication after transient deoxynucleotide depletion, and all display synthetic effects when also carrying mutations affecting the polα-primase and RPA DNA replication complexes, suggesting a role for Bmh proteins in DNA replication stress response. Finally, the bmh1-169 bmh2Δ and bmh1-170 bmh2Δ mutants show increased rates of spontaneous gross chromosomal rearrangements, indicating that Bmh proteins are required to suppress genome instability.

Understanding the functions of BRCA1 in the DNA-damage response

2009 ◽  
Vol 37 (3) ◽  
pp. 597-604 ◽  
Author(s):  
Maximina H. Yun ◽  
Kevin Hiom
Keyword(s):  
Dna Damage ◽  
Early Onset ◽  
Genome Stability ◽  
Chromosomal Rearrangements ◽  
Dna Lesions ◽  
Cell Cycle Checkpoints ◽  
Dna Breaks ◽  
Gross Chromosomal Rearrangements ◽  
Damage Response ◽  
Regulation Of Cell Cycle

Inheritance of a mutation in BRCA1 (breast cancer 1 early-onset) results in predisposition to early-onset breast and ovarian cancer. Tumours in these individuals arise after somatic mutation or loss of the wild-type allele. Loss of BRCA1 function leads to a profound increase in genomic instability involving the accumulation of mutations, DNA breaks and gross chromosomal rearrangements. Accordingly, BRCA1 has been implicated as an important factor involved in both the repair of DNA lesions and in the regulation of cell-cycle checkpoints in response to DNA damage. However, the molecular mechanism through which BRCA1 functions to preserve genome stability remains unclear. In the present article, we examine the different ways in which BRCA1 might influence the repair of DNA damage and the preservation of genome integrity, taking into account what is currently known about its interactions with other proteins, its biochemical activity and its nuclear localization.

scholarly journals Inactivation of folylpolyglutamate synthetase Met7 results in genome instability driven by an increased dUTP/dTTP ratio

2019 ◽  
Author(s):  
Tobias T Schmidt ◽  
Sushma Sharma ◽  
Gloria X Reyes ◽  
Anna Kolodziejczak ◽  
Tina Wagner ◽  
...  
Keyword(s):  
Dna Damage ◽  
Genome Stability ◽  
Nuclear Dna ◽  
Genome Instability ◽  
Chromosomal Rearrangements ◽  
Dna Integrity ◽  
Folylpolyglutamate Synthetase ◽  
Cell Extracts ◽  
Gross Chromosomal Rearrangements ◽  
The Stability

Abstract The accumulation of mutations is frequently associated with alterations in gene function leading to the onset of diseases, including cancer. Aiming to find novel genes that contribute to the stability of the genome, we screened the Saccharomyces cerevisiae deletion collection for increased mutator phenotypes. Among the identified genes, we discovered MET7, which encodes folylpolyglutamate synthetase (FPGS), an enzyme that facilitates several folate-dependent reactions including the synthesis of purines, thymidylate (dTMP) and DNA methylation. Here, we found that Met7-deficient strains show elevated mutation rates, but also increased levels of endogenous DNA damage resulting in gross chromosomal rearrangements (GCRs). Quantification of deoxyribonucleotide (dNTP) pools in cell extracts from met7Δ mutant revealed reductions in dTTP and dGTP that cause a constitutively active DNA damage checkpoint. In addition, we found that the absence of Met7 leads to dUTP accumulation, at levels that allowed its detection in yeast extracts for the first time. Consequently, a high dUTP/dTTP ratio promotes uracil incorporation into DNA, followed by futile repair cycles that compromise both mitochondrial and nuclear DNA integrity. In summary, this work highlights the importance of folate polyglutamylation in the maintenance of nucleotide homeostasis and genome stability.

scholarly journals Gross chromosomal rearrangements and genetic exchange between nonhomologous chromosomes following BRCA2 inactivation

2000 ◽  
Vol 14 (11) ◽  
pp. 1400-1406 ◽  
Author(s):  
Veronica P.C.C. Yu ◽  
Michael Koehler ◽  
Claus Steinlein ◽  
Michael Schmid ◽  
Leslyn A. Hanakahi ◽  
...  
Keyword(s):  
Dna Damage ◽  
Genetic Instability ◽  
Genomic Dna ◽  
Chromosomal Rearrangements ◽  
Chromosome Breakage ◽  
Genetic Exchange ◽  
Dna Breaks ◽  
Gross Chromosomal Rearrangements ◽  
Essential Function ◽  
Mechanistic Basis

Cancer-causing mutations often arise from gross chromosomal rearrangements (GCRs) such as translocations, which involve genetic exchange between nonhomologous chromosomes. Here we show that murineBrca2 has an essential function in suppressing GCR formation after chromosome breakage. Cells that harbor truncated Brca2spontaneously incur GCRs and genomic DNA breaks during division. They exhibit hypersensitivity to DNA damage by interstrand cross-linkers, which even at low doses trigger aberrant genetic exchange between nonhomologous chromosomes. Therefore, genetic instability in Brca2-deficient cells results from the mutagenic processing of spontaneous or induced DNA damage into gross chromosomal rearrangements, providing a mechanistic basis for cancer predisposition.

scholarly journals Cdc28/Cdk1 positively and negatively affects genome stability in S. cerevisiae

2009 ◽  
Vol 185 (3) ◽  
pp. 423-437 ◽  
Author(s):  
Jorrit M. Enserink ◽  
Hans Hombauer ◽  
Meng-Er Huang ◽  
Richard D. Kolodner
Keyword(s):  
Dna Damage ◽  
Cell Viability ◽  
Genome Stability ◽  
Chromosomal Rearrangements ◽  
Genetic Network ◽  
Telomere Maintenance ◽  
Mitotic Catastrophe ◽  
Chemical Genetic ◽  
Replication Arrest ◽  
Gross Chromosomal Rearrangements

We studied the function of the cyclin-dependent kinase Cdc28 (Cdk1) in the DNA damage response and maintenance of genome stability using Saccharomyces cerevisiae. Reduced Cdc28 activity sensitizes cells to chronic DNA damage, but Cdc28 is not required for cell viability upon acute exposure to DNA-damaging agents. Cdc28 is also not required for activation of the DNA damage and replication checkpoints. Chemical–genetic analysis reveals that CDC28 functions in an extensive network of pathways involved in maintenance of genome stability, including homologous recombination, sister chromatid cohesion, the spindle checkpoint, postreplication repair, and telomere maintenance. In addition, Cdc28 and Mre11 appear to cooperate to prevent mitotic catastrophe after DNA replication arrest. We show that reduced Cdc28 activity results in suppression of gross chromosomal rearrangements (GCRs), indicating that Cdc28 is required for formation or recovery of GCRs. Thus, we conclude that Cdc28 functions in a genetic network that supports cell viability during DNA damage while promoting the formation of GCRs.

scholarly journals Oncogene homologue Sch9 promotes age-dependent mutations by a superoxide and Rev1/Polζ-dependent mechanism

2009 ◽  
Vol 186 (4) ◽  
pp. 509-523 ◽  
Author(s):  
Federica Madia ◽  
Min Wei ◽  
Valerie Yuan ◽  
Jia Hu ◽  
Cristina Gattazzo ◽  
...  
Keyword(s):  
Dna Damage ◽  
Chromosomal Rearrangements ◽  
Point Mutations ◽  
Mitochondrial Superoxide ◽  
Gross Chromosomal Rearrangements ◽  
Age Dependent ◽  
Induced Mutagenesis ◽  
Major Increase ◽  
Oncogene Homologue ◽  
Mitochondrial Superoxide Dismutase

Oncogenes contribute to tumorigenesis by promoting growth and inhibiting apoptosis. Here we examine the function of Sch9, the Saccharomyces cerevisiae homologue of the mammalian Akt and S6 kinase, in DNA damage and genomic instability during aging in nondividing cells. Attenuation of age-dependent increases in base substitutions, small DNA insertions/deletions, and gross chromosomal rearrangements (GCRs) in sch9Δ mutants is associated with increased mitochondrial superoxide dismutase (MnSOD) expression, decreased DNA oxidation, reduced REV1 expression and translesion synthesis, and elevated resistance to oxidative stress-induced mutagenesis. Deletion of REV1, the lack of components of the error-prone Polζ, or the overexpression of SOD1 or SOD2 is sufficient to reduce age-dependent point mutations in SCH9 overexpressors, but REV1 deficiency causes a major increase in GCRs. These results suggest that the proto-oncogene homologue Sch9 promotes the accumulation of superoxide-dependent DNA damage in nondividing cells, which induces error-prone DNA repair that generates point mutations to avoid GCRs and cell death during the first round of replication.

scholarly journals Defects in DNA Lesion Bypass Lead to Spontaneous Chromosomal Rearrangements and Increased Cell Death

2009 ◽  
Vol 9 (2) ◽  
pp. 315-324 ◽  
Author(s):  
Kristina H. Schmidt ◽  
Emilie B. Viebranz ◽  
Lorena B. Harris ◽  
Hamed Mirzaei-Souderjani ◽  
Salahuddin Syed ◽  
...  
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Genetic Interaction ◽  
Chromosomal Rearrangements ◽  
Point Mutations ◽  
Normal Growth ◽  
Dna Helicase ◽  
Dna Damage Checkpoint ◽  
Lesion Bypass

ABSTRACT Rev3 polymerase and Mph1 DNA helicase participate in error-prone and error-free pathways, respectively, for the bypassing of template lesions during DNA replication. Here we have investigated the role of these pathways and their genetic interaction with recombination factors, other nonreplicative DNA helicases, and DNA damage checkpoint components in the maintenance of genome stability, viability, and sensitivity to the DNA-damaging agent methyl methanesulfonate (MMS). We find that cells lacking Rev3 and Mph1 exhibit a synergistic, Srs2-dependent increase in the rate of accumulating spontaneous, gross chromosomal rearrangements, suggesting that the suppression of point mutations by deletion of REV3 may lead to chromosomal rearrangements. While mph1Δ is epistatic to homologous recombination (HR) genes, both Rad51 and Rad52, but not Rad59, are required for normal growth of the rev3Δ mutant and are essential for survival of rev3Δ cells during exposure to MMS, indicating that Mph1 acts in a Rad51-dependent, Rad59-independent subpathway of HR-mediated lesion bypass. Deletion of MPH1 helicase leads to synergistic DNA damage sensitivity increases in cells with chl1Δ or rrm3Δ helicase mutations, whereas mph1Δ is hypostatic to sgs1Δ. Previously reported slow growth of mph1Δ srs2Δ cells is accompanied by G2/M arrest and fully suppressed by disruption of the Mec3-dependent DNA damage checkpoint. We propose a model for replication fork rescue mediated by translesion DNA synthesis and homologous recombination that integrates the role of Mph1 in unwinding D loops and its genetic interaction with Rev3 and Srs2-regulated pathways in the suppression of spontaneous genome rearrangements and in mutation avoidance.

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