DNA2 drives processing and restart of reversed replication forks in human cells

Saravanabhavan Thangavel, Matteo Berti, Maryna Levikova, Cosimo Pinto, Shivasankari Gomathinayagam, Marko Vujanovic, Ralph Zellweger, Hayley Moore, Eu Han Lee, Eric A. Hendrickson, Petr Cejka, Sheila Stewart, Massimo Lopes, Alessandro Vindigni

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

Accurate processing of stalled or damaged DNA replication forks is paramount to genomic integrity and recent work points to replication fork reversal and restart as a central mechanism to ensuring high-fidelity DNA replication. Here, we identify a novel DNA2- and WRN-dependent mechanism of reversed replication fork processing and restart after prolonged genotoxic stress. The human DNA2 nuclease and WRN ATPase activities functionally interact to degrade reversed replication forks with a 5'-to-3' polarity and promote replication restart, thus preventing aberrant processing of unresolved replication intermediates. Unexpectedly, EXO1, MRE11, and CtIP are not involved in the same mechanism of reversed fork processing, whereas human RECQ1 limits DNA2 activity by preventing extensive nascent strand degradation. RAD51 depletion antagonizes this mechanism, presumably by preventing reversed fork formation. These studies define a new mechanism for maintaining genome integrity tightly controlled by specific nucleolytic activities and central homologous recombination factors.

Original languageEnglish (US)
Pages (from-to)545-562
Number of pages18
JournalJournal of Cell Biology
Volume208
Issue number5
DOIs
StatePublished - Jan 1 2015

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DNA Replication
Homologous Recombination
DNA Damage
Adenosine Triphosphatases
Genome

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Thangavel, S., Berti, M., Levikova, M., Pinto, C., Gomathinayagam, S., Vujanovic, M., ... Vindigni, A. (2015). DNA2 drives processing and restart of reversed replication forks in human cells. Journal of Cell Biology, 208(5), 545-562. https://doi.org/10.1083/jcb.201406100

DNA2 drives processing and restart of reversed replication forks in human cells. / Thangavel, Saravanabhavan; Berti, Matteo; Levikova, Maryna; Pinto, Cosimo; Gomathinayagam, Shivasankari; Vujanovic, Marko; Zellweger, Ralph; Moore, Hayley; Lee, Eu Han; Hendrickson, Eric A.; Cejka, Petr; Stewart, Sheila; Lopes, Massimo; Vindigni, Alessandro.

In: Journal of Cell Biology, Vol. 208, No. 5, 01.01.2015, p. 545-562.

Research output: Contribution to journalArticle

Thangavel, S, Berti, M, Levikova, M, Pinto, C, Gomathinayagam, S, Vujanovic, M, Zellweger, R, Moore, H, Lee, EH, Hendrickson, EA, Cejka, P, Stewart, S, Lopes, M & Vindigni, A 2015, 'DNA2 drives processing and restart of reversed replication forks in human cells', Journal of Cell Biology, vol. 208, no. 5, pp. 545-562. https://doi.org/10.1083/jcb.201406100
Thangavel S, Berti M, Levikova M, Pinto C, Gomathinayagam S, Vujanovic M et al. DNA2 drives processing and restart of reversed replication forks in human cells. Journal of Cell Biology. 2015 Jan 1;208(5):545-562. https://doi.org/10.1083/jcb.201406100
Thangavel, Saravanabhavan ; Berti, Matteo ; Levikova, Maryna ; Pinto, Cosimo ; Gomathinayagam, Shivasankari ; Vujanovic, Marko ; Zellweger, Ralph ; Moore, Hayley ; Lee, Eu Han ; Hendrickson, Eric A. ; Cejka, Petr ; Stewart, Sheila ; Lopes, Massimo ; Vindigni, Alessandro. / DNA2 drives processing and restart of reversed replication forks in human cells. In: Journal of Cell Biology. 2015 ; Vol. 208, No. 5. pp. 545-562.
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