Regulation of proliferating cell nuclear antigen ubiquitination in mammalian cells

Niimi, Atsuko, Brown, Stephanie, Sabbioneda, Simone, Kannouche, Patricia L., Scott, Andrew, Yasui, Akira, Green, Catherine M. and Lehmann, Alan R. (2008) Regulation of proliferating cell nuclear antigen ubiquitination in mammalian cells. Proceedings of the National Academy of Sciences, 105 (42). pp. 16125-16130. ISSN 0027-8424

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After exposure to DNA-damaging agents that block the progress of the replication fork, monoubiquitination of proliferating cell nuclear antigen (PCNA) mediates the switch from replicative to translesion synthesis DNA polymerases. We show that in human cells, PCNA is monoubiquitinated in response to methyl methanesulfonate and mitomycin C, as well as UV light, albeit with different kinetics, but not in response to bleomycin or camptothecin. Cyclobutane pyrimidine dimers are responsible for most of the PCNA ubiquitination events after UV-irradiation. Failure to ubiquitinate PCNA results in substantial sensitivity to UV and methyl methanesulfonate, but not to camptothecin or bleomycin. PCNA ubiquitination depends on Replication Protein A (RPA), but is independent of ATR-mediated checkpoint activation. After UV-irradiation, there is a temporal correlation between the disappearance of the deubiquitinating enzyme USP1 and the presence of PCNA ubiquitination, but this correlation was not found after chemical mutagen treatment. By using cells expressing photolyases, we are able to remove the UV lesions, and we show that PCNA ubiquitination persists for many hours after the damage has been removed. We present a model of translesion synthesis behind the replication fork to explain the persistence of ubiquitinated PCNA.

Item Type: Article
Additional Information: In Endnotes Ref: GDSC263
Keywords: DNA replication, UV damage, translesion synthesis
Schools and Departments: School of Life Sciences > Biochemistry
School of Life Sciences > Sussex Centre for Genome Damage and Stability
Subjects: Q Science > QD Chemistry > QD0241 Organic chemistry > QD0415 Biochemistry
Q Science > QL Zoology
R Medicine > R Medicine (General) > R855 Medical technology
Depositing User: Gee Wheatley
Date Deposited: 23 Feb 2009
Last Modified: 03 Jul 2019 00:07
Google Scholar:52 Citations

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