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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 ISSN 1932-6203.

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Official URL: http://dx.doi.org/10.1371/journal.pone.0047091

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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
Subjects: Q Science > QK Botany
Divisions: Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology
Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School
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:
DateEvent
2012Published
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 (Creative Commons)
Date of first compliant deposit: 23 December 2015
Date of first compliant Open Access: 23 December 2015
Funder: Marie Curie Cancer Care

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