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DNA polymerase α (swi7) and the flap endonuclease fen1 (rad2) act together in the s-phase alkylation damage response in S. pombe
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Koulintchenko, Milana, Vengrova, Sonya, Eydmann, Trevor, Arumugam, Prakash and Dalgaard, Jacob Z. (2012) DNA polymerase α (swi7) and the flap endonuclease fen1 (rad2) act together in the s-phase alkylation damage response in S. pombe. PLoS One, Vol.7 (No.10). e47091. doi:10.1371/journal.pone.0047091
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Official URL: http://dx.doi.org/10.1371/journal.pone.0047091
Abstract
Polymerase α is an essential enzyme mainly mediating Okazaki fragment synthesis during lagging strand replication. A specific point mutation in Schizosaccharomyces pombe polymerase α named swi7-1, abolishes imprinting required for mating-type switching. Here we investigate whether this mutation confers any genome-wide defects. We show that the swi7-1 mutation renders cells hypersensitive to the DNA damaging agents methyl methansulfonate (MMS), hydroxyurea (HU) and UV and incapacitates activation of the intra-S checkpoint in response to DNA damage. In addition we show that, in the swi7-1 background, cells are characterized by an elevated level of repair foci and recombination, indicative of increased genetic instability. Furthermore, we detect novel Swi1-, -Swi3- and Pol α- dependent alkylation damage repair intermediates with mobility on 2D-gel that suggests presence of single-stranded regions. Genetic interaction studies showed that the flap endonuclease Fen1 works in the same pathway as Pol α in terms of alkylation damage response. Fen1 was also required for formation of alkylation- damage specific repair intermediates. We propose a model to explain how Pol α, Swi1, Swi3 and Fen1 might act together to detect and repair alkylation damage during S-phase.
| Item Type: | Journal Article | ||||
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| Subjects: | Q Science > QK Botany | ||||
| Divisions: | Faculty of Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology Faculty of Science > Life Sciences (2010- ) Faculty of Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | DNA polymerases, Schizosaccharomyces pombe -- Physiology, Mutation (Biology) | ||||
| Journal or Publication Title: | PLoS One | ||||
| Publisher: | Public Library of Science | ||||
| ISSN: | 1932-6203 | ||||
| Official Date: | 2012 | ||||
| Dates: |
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| Volume: | Vol.7 | ||||
| Number: | No.10 | ||||
| Page Range: | e47091 | ||||
| DOI: | 10.1371/journal.pone.0047091 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Open Access | ||||
| Funder: | Marie Curie Cancer Care |
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