TCR in yeast Telomeres

Published: 6 October 2021| Version 1 | DOI: 10.17632/j9869tptct.1
Contributors:
,
Audrey Paillé,
,

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Raw Data associated to the Guintini et al, TCR in yeast Telomeres, 2021 article T4endonucleaseV results : NER in the Y’-element telomeres. WT and rad26Δ yeast were irradiated at 180 J/m2 and harvested at the indicated times in hours (hrs). Isolated DNA from non-irradiated (−UV) and irradiated cells (0 to 4 hrs) were digested with HindIII, EcoRI and XhoI, and mock treated or treated with T4 endo-V (T4-V), denoted by − and +, respectively. After separation of the DNA fragments in 1% alkaline agarose-gels and blotting, the filter membranes were hybridized with single-stranded, strand-specific oligonucleotides. The band signals correspond to the average of all Y’-elements in the cell. Quantification of CPDs in the ~4.7 kb HindIII/EcoRI group of bands formed by the 4.4, 4.7 and 5.0 kb fragments (brackets). Quantification of CPDs in the ~3.0 kb HindIII fragment, made of ~2.7 kb of Y’-element sequences and variable lengths (average 0.3 kb) of telomere repeats. Measurements were taken of the broad bands (brackets). Quantification of CPDs in the 1.3 kb XhoI fragment, made of ~1 kb of Y’-element sequences and telomere repeats. Data are from quantification of phosphor images for the C-rich and G-rich strands of WT and rad26. Primer Extension results : Repair of pyrimidine dimers in the Tel15L X-element of WT, rad26, sir2 and sir2rad26 cells of Crich and G-rich strand. Yeast were UV irradiated at 180 J/m2 and harvested after 0, 0.5, 1, 2 and 4 hours (hrs). Isolated DNA from non-irradiated (U) and irradiated cells (0 to 4 hrs) were digested with RsaI or HhaI and used as template for TaqI polymerase primer-extension. Repair of PDs was assessed by the extension of primers 'a', 'b', ‘c’ and ‘d’.

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DNA Repair

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