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Effect of proliferating cell nuclear antigen ubiquitination and chromatin structure on the dynamic properties of the Y-family DNA polymerases

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posted on 2023-06-07, 14:58 authored by Simone Sabbioneda, Audrey M Gourdin, Catherine M Green, Angelika Zotter, Giuseppina Giglia-Mari, Adriaan Houtsmuller, Wim Vermeulen, Alan LehmannAlan Lehmann
Y-family DNA polymerases carry out translesion synthesis past damaged DNA. DNA polymerases (pol) ? and ? are usually uniformly distributed through the nucleus but accumulate in replication foci during S phase. DNA-damaging treatments result in an increase in S phase cells containing polymerase foci. Using photobleaching techniques, we show that pol? is highly mobile in human fibroblasts. Even when localized in replication foci, it is only transiently immobilized. Although ubiquitination of proliferating cell nuclear antigen (PCNA) is not required for the localization of pol? in foci, it results in an increased residence time in foci. pol? is even more mobile than pol?, both when uniformly distributed and when localized in foci. Kinetic modeling suggests that both pol? and pol? diffuse through the cell but that they are transiently immobilized for ~150 ms, with a larger proportion of pol? than pol? immobilized at any time. Treatment of cells with DRAQ5, which results in temporary opening of the chromatin structure, causes a dramatic immobilization of pol? but not pol?. Our data are consistent with a model in which the polymerases are transiently probing the DNA/chromatin. When DNA is exposed at replication forks, the polymerase residence times increase, and this is further facilitated by the ubiquitination of PCNA.

History

Publication status

  • Published

File Version

  • Published version

Journal

Molecular Biology of the Cell

ISSN

1939-4586

Publisher

American Society for Cell Biology

Issue

12

Volume

19

Page range

5193-5202

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Notes

Endnotes GDSC282 PMCID: PMC2592664

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2009-03-25

First Open Access (FOA) Date

2018-03-21

First Compliant Deposit (FCD) Date

2018-03-21

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