Abstract: DNA-protein cross-links (DPCs) are extremely bulky adducts that interfere with replication. In human cells, they are processed by SPRTN, a protease activated by DNA polymerases stuck at DPCs. We have recently proposed the mechanism of the interaction of DNA polymerases with DPCs, involving a clash of protein surfaces followed by the distortion of the cross-linked protein. Here, we used a model DPC, located in the single-stranded template, the template strand of double-stranded DNA, or the displaced strand, to study the eukaryotic translesion DNA polymerases zeta (POL zeta), iota (POL iota) and eta (POL eta). POL iota demonstrated poor synthesis on the DPC-containing substrates. POL zeta and POL eta paused at sites dictated by the footprints of the polymerase and the cross-linked protein. Beyond that, POL zeta was able to elongate the primer to the cross-link site when a DPC was in the template. Surprisingly, POL eta was not only able to reach the cross-link site but also incorporated 1-2 nucleotides past it, which makes POL eta the most efficient DNA polymerase on DPC-containing substrates. However, a DPC in the displaced strand was an insurmountable obstacle for all polymerases, which stalled several nucleotides before the cross-link site. Overall, the behavior of translesion polymerases agrees with the model of protein clash and distortion described above.

Cite: Yudkina A.V. , Shilkin E.S. , Makarova A.V. , Zharkov D.O.
Stalling of Eukaryotic Translesion DNA Polymerases at DNA-Protein Cross-Links
Genes. 2022. V.13. N2. 166 . DOI: 10.3390/genes13020166 WOS Scopus OpenAlex

Identifiers:

Web of science:	WOS:000769507400001
Scopus:	2-s2.0-85125349182
OpenAlex:	W4205931361

Citing:

DB	Citing
Web of science	9
Scopus	6
OpenAlex	8

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