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| Status |
Public on Mar 26, 2022 |
| Title |
Ribodysgenesis: sudden genome instability in the yeast Saccharomyces cerevisiae arising from RNase H2 cleavage at genomic-embedded ribonucleotides |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Other
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| Summary |
Yeast Saccharomyces cerevisiae has been widely used as a model system for studying genome instability. Here, heterozygous S. cerevisiae zygotes were generated to determine the genomic alterations induced by sudden introduction of active RNase H2. In combination of a custom SNP microarray, the patterns of chromosomal instability could be explored at a whole genome level.
Ribonucleotides can be incorporated into DNA during replication by the replicative DNA polymerases. These aberrant DNA subunits are efficiently recognized and removed by Ribonucleotide Excision Repair, which is initiated by the heterotrimeric enzyme RNase H2. While RNase H2 is essential in higher eukaryotes, the yeast Saccharomyces cerevisiae can survive without RNase H2 enzyme, although the genome undergoes mutation, recombination and other genome instability events at an increased rate. Although RNase H2 can be considered as a protector of the genome from the deleterious events that can ensue from recognition and removal of embedded ribonucleotides, under conditions of high ribonucleotide incorporation and retention in the genome in a RNase H2-negative strain, sudden introduction of active RNase H2 causes massive DNA breaks and genome instability in a condition which we term “ribodysgenesis”. The DNA breaks and genome instability arise solely from RNase H2 cleavage directed to the ribonucleotide-containing genome. Survivors of ribodysgenesis have massive loss of heterozygosity events stemming from recombinogenic lesions on the ribonucleotide-containing DNA, with increases of over 1000X from wild-type. DNA breaks are produced over one to two divisions and subsequently cells adapt to RNase H2 and ribonucleotides in the genome and grow with normal levels of genome instability.
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| Overall design |
Zygotes were grown on on solid YPD medium or in liquid YPD medium at 30°C. Genomic DNA from independent zygotes was extracted and analyzed by the whole-genome microarrays.
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| Contributor(s) |
Zheng D, Petes TD |
| Citation missing |
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| Submission date |
Mar 23, 2022 |
| Last update date |
Mar 28, 2022 |
| Contact name |
Daoqiong Zheng |
| E-mail(s) |
zhengdaoqiong@163.com
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| Phone |
86 571 88206636
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| Organization name |
College of Life Sciences, Zhejiang University
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| Department |
Institute of Microbiology
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| Street address |
866 Yuhangtang Road
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| City |
Hangzhou |
| State/province |
Zhejiang |
| ZIP/Postal code |
310058 |
| Country |
China |
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| Platforms (1) |
| GPL21553 |
Agilent-031671 Chr4 RA V2 027289 (SPOT_ID version) |
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| Samples (10)
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| GSM5968379 |
Cross of MD893-1 with LSY3454-2 Zygote1 |
| GSM5968380 |
Cross of MD893-1 with LSY3454-2 Zygote2 |
| GSM5968381 |
Cross of MD893-1 with LSY3454-2 Zygote3 |
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| Relations |
| BioProject |
PRJNA819077 |
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