Structure of an archaeal B‐family DNA polymerase in complex with a chemically modified nucleotide

Archaeal B‐family DNA polymerases (DNA pols) are the driving force of cutting edge biotechnological applications like next‐generation sequencing. The acceptance of chemically modified nucleotides by the DNA pols is key to these technologies. By now, no structural data are available for these DNA pols in complex with modified substrates which could build the basis for understanding interactions between the enzyme and the chemical modification and for further development of next generation nucleotides. For the first time we crystallized an exonuclease deficient variant of B‐family KOD DNA pol wild‐type with a modified nucleotide in a closed, ternary complex. We also crystalized the A‐family DNA pol KlenTaq with the same nucleotide. The reported structural data elucidates how the protein and the DNA modulate two distinct conformations of the modification in the A‐ and B‐family DNA pols and how these influence the processing of the modified nucleotide. Overall, this study provides first insights on how B‐family DNA pols interplay with relevant modified substrates.

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