TCR in yeast Telomeres

Raw Data associated to the Guitini et al (2022) Transcription of ncRNAs promotes repair of UV induced DNA lesions in Saccharomyces cerevisiae subtelomeres. Plos Genetics.

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’.