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Supplemental Material for Stepchenkova et al., 2021

journal contribution
posted on 08.04.2021, 18:51 by Elena I. Stepchenkova, Anna S. Zhuk, Jian Cui, Elena R. Tarakhovskaya, Stephanie R. Barbari, Polina V. Shcherbakova, Dmitrii E. Polev, Roman Fedorov, Eugenia Poliakov, Igor B. Rogozin, Artem G. Lada, Youri I Pavlov
Supplemental materials include 11 Suppl figures - pdf-files,
6 Suppl Tables, 4 pdf and 2 excel files
Suppl Results, 2 pdf files.

Supplemental Results 1. Problems with the construction of haploid yeast strains with DNA polymerase alleles reducing fitness.

Supplemental Results 2. Estimation of the probabilities of recurrent mutations in the same gene.

Supplemental Table 1. Mutator effect of pol2rc-ΔN is reduced 60% in strain without the catalytic subunit of pol ζ, Rev3.

Supplemental Table 2. Differences in mutant frequencies with or without UV-light irradiation in pol2rc-ΔN vs. wild-type strains.

Supplemental Table 3. The pol2rc-ΔN elevated chromosome instability.

Supplemental Table 4. (Excel file) Summary of mutations found in the course of genomic sequencing of the pol2rc-ΔN segregants.

Supplementary Table 5. (Excel file) Ancestry of mutations in independent clones of the pol2rc-ΔNsegregants.

Supplemental Table 6. Types of mutations found in genomes of pol2rc-ΔΝ strains.

Supplemental Figure 1. Creation of haploid pol2rc-ΔN strains.

Supplemental Figure 2. The absence of DPB4 does not affect the slow growth phenotype of pol2-ΔNmutants.

Supplemental Figure 3. Outline of genetic analysis of diploids double heterozygous for the complete deletion of POL2 gene and pol2rc-ΔN.

Supplemental Figure 4. Peculiarities in the genomic regions of pol2rc-ΔN strains relevant to the construction method.

Supplemental Figure 5. High proportion of cells with abnormal morphology and nuclear material in pol2rc-ΔN cells.

Supplemental Figure 6. Strains with pol2rc-ΔN grow slower than wild-type but are not temperature-sensitive or cold-sensitive.

Supplemental Figure 7. Higher proportion of dead methylene blue-stained cells in cultures of pol2rc-ΔN strains in comparison to wild-type strains.

Supplemental Figure 8. Many cells with pol2rc-ΔN have difficulty to start dividing.

Supplemental Figure 9. Schematic of analysis to find if single cdc28 mutations can confer a growth advantage to pol2rc-ΔN strains.

Supplementary Figure 10. Rescue of growth defects and sensitivity to drugs of pol2rc-ΔN by cdc28 mutations.

Supplementary Figure 11. Multiple alignment of CMGC/CDK/CDC2 protein kinase orthologs.


History

Article title

Compensation for the absence of the catalytically active half of DNA polymerase ε in yeast by positively selected mutations in CDC28