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The histone methyltransferase SET8 is required for S-phase progression
University of Copenhagen, Biotech Research and Innovation Centre. (Claus Storgaard Sorensen)
Stockholm University, Faculty of Science, Department of Genetics, Microbiology and Toxicology. (Thomas Helleday)
University of Southern Denmark, Centre for Epigenetics, Department of Biochemistry and Molecular Biology.
University of Copenhagen, Biotech Research and Innovation Centre.
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2007 (English)In: Journal of Cell Biology, ISSN 0021-9525, E-ISSN 1540-8140, Vol. 179, no 7, 1337-1345 p.Article in journal (Refereed) Published
Abstract [en]

Chromatin structure and function is influenced by histone posttranslational modifications. SET8 (also known as PR-Set7 and SETD8) is a histone methyltransferase that monomethylates histone H4-K20. However, a function for SET8 in mammalian cell proliferation has not been determined. We show that small interfering RNA inhibition of SET8 expression leads to decreased cell proliferation and accumulation of cells in S phase. This is accompanied by DNA double-strand break (DSB) induction and recruitment of the DNA repair proteins replication protein A, Rad51, and 53BP1 to damaged regions. SET8 depletion causes DNA damage specifically during replication, which induces a Chk1-mediated S-phase checkpoint. Furthermore, we find that SET8 interacts with proliferating cell nuclear antigen through a conserved motif, and SET8 is required for DNA replication fork progression. Finally, codepletion of Rad51, an improtant homologous recombination repair protein, abrogates the DNA damage after SET8 depletion. Overall, we show that SET8 is essential for genomic stability in mammalian cells and that decreased expression of SET8 results in DNA damage and Chk1-dependent S-phase arrest.

Place, publisher, year, edition, pages
2007. Vol. 179, no 7, 1337-1345 p.
National Category
Natural Sciences
URN: urn:nbn:se:su:diva-56695DOI: 10.1083/jcb.200706150OAI: diva2:412231
Available from: 2011-04-21 Created: 2011-04-21 Last updated: 2011-04-27Bibliographically approved
In thesis
1. Replication Fork Stability in Mammalian Cells
Open this publication in new window or tab >>Replication Fork Stability in Mammalian Cells
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Maintaining replication fork integrity is vital to preserve genomic stability and avoid cancer. Physical DNA damage and altered nucleotide or protein pools represent replication obstacles, generating replicative stress. Numerous cellular responses have evolved to ensure faithful DNA replication despite such challenges. Understanding those responses is essential to understand and prevent or treat replication-associated diseases, such as cancer.

Re-priming is a mechanism to allow resumption of DNA synthesis past a fork-stalling lesion. This was recently suggested in yeast and explains the formation of gaps during DNA replication on damaged DNA. Using a combination of assays, we indicate the existence of re-priming also in human cells following UV irradiation.

The gap left behind a re-primed fork must be stabilised to avoid replication-associated collapse. Our results show that the checkpoint signalling protein CHK1 is dispensable for stabilisation of replication forks after UV irradiation, despite its role in replication fork progression on UV-damaged DNA. It is not known what proteins are necessary for collapse of an unsealed gap or a stalled fork. We exclude one, previously suggested, endonuclease from this mechanism in UV-irradiated human fibroblasts. We also show that focus formation of repair protein RAD51 is not necessarily associated with cellular sensitivity to agents inducing replicative stress, in rad51d CHO mutant cells.

Multiple factors are required for replication fork stability, also under unperturbed conditions. We identify the histone methyltransferase SET8 as an important player in the maintenance of replication fork stability. SET8 is required for replication fork progression, and depletion of SET8 led to the formation of replication-associated DNA damage.

In summary, our results increase the knowledge about mechanisms and signalling at replication forks in unperturbed cells and after induction of replicative stress.

Place, publisher, year, edition, pages
Stockholm: Department of Genetics, Microbiology and Toxicology, Stockholm University, 2011. 77 p.
replication fork progression, replication fork stability, re-priming, DNA damage signalling
National Category
Natural Sciences
Research subject
Molecular Genetics
urn:nbn:se:su:diva-56697 (URN)978-91-7447-270-7 (ISBN)
Public defence
2011-05-26, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 16 B, Stockholm, 10:00 (English)
At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 1: Submitted. Paper 2: Submitted. Paper 3: Manuscript. Paper 5: Submitted.Available from: 2011-05-04 Created: 2011-04-21 Last updated: 2011-06-21Bibliographically approved

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Elvers, IngegerdHelleday, Thomas
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