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. Author manuscript; available in PMC: 2020 Dec 2.
Published in final edited form as: Cell Rep. 2020 Nov 17;33(7):108401. doi: 10.1016/j.celrep.2020.108401

Figure 3. Mapping Atypical TA Photoproducts in UV-Irradiated DNA and Cells.

Figure 3.

(A) Design of DNA oligonucleotide containing a stretch of five thymine-adenine (TA) sequences (underlined), but no dipyrimidine sequences.

(B) Analysis of UV lesions by denaturing gel electrophoresis. Double-stranded DNA oligos were irradiated with increasing doses of UVC light (0.86–25.7 kJ/m2) and cleaved by UV DNA endonuclease (UVDE). The locations of the different TA photoproducts in the DNA sequence are indicated (TA 1–5), based on size standards in the leftmost lane.

(C) Quantification of UVDE-cleaved TA photoproducts induced by different doses of UVC light.

(D) Chemical structure of TA photoproduct.

(E) UVDE-seq method for mapping non-CPD UV photoproducts. CPD lesions are removed by photoreactivation with purified CPD photolyase, and the remaining UV photoproducts are cleaved with UVDE.

(F) UVDE-seq reads are enriched at dipyrimidine sequences and TA dinucleotides immediately following irradiation of a rad16Δ mutant with 600 J/m2 UVC light, consistent with UV-induced formation of 6–4PPs and TA photoproducts. “No UV” sample is included as a control.