Damage-induced DNA replication stalling relies on MAPK-activated protein kinase 2 activity

TY - JOUR

T1 - Damage-induced DNA replication stalling relies on MAPK-activated protein kinase 2 activity

AU - Köpper, Frederik

AU - Bierwirth, Cathrin

AU - Schön, Margarete

AU - Kunze, Meike

AU - Elvers, Ingegerd

AU - Kranz, Dominique

AU - Saini, Priyanka

AU - Menon, Manoj B

AU - Walter, David

AU - Sørensen, Claus Storgaard

AU - Gaestel, Matthias

AU - Helleday, Thomas

AU - Schön, Michael P

AU - Dobbelstein, Matthias

PY - 2013/10/15

Y1 - 2013/10/15

N2 - DNA damage can obstruct replication forks, resulting in replicative stress. By siRNA screening, we identified kinases involved in the accumulation of phosphohistone 2AX (γH2AX) upon UV irradiation-induced replication stress. Surprisingly, the strongest reduction of phosphohistone 2AX followed knockdown of the MAP kinase-activated protein kinase 2 (MK2), a kinase currently implicated in p38 stress signaling and G2 arrest. Depletion or inhibition of MK2 also protected cells from DNA damage-induced cell death, and mice deficient for MK2 displayed decreased apoptosis in the skin upon UV irradiation. Moreover, MK2 activity was required for damage response, accumulation of ssDNA, and decreased survival when cells were treated with the nucleoside analogue gemcitabine or when the checkpoint kinase Chk1 was antagonized. By using DNA fiber assays, we found that MK2 inhibition or knockdown rescued DNA replication impaired by gemcitabine or by Chk1 inhibition. This rescue strictly depended on translesion DNA polymerases. In conclusion, instead of being an unavoidable consequence of DNA damage, alterations of replication speed and origin firing depend on MK2-mediated signaling.

AB - DNA damage can obstruct replication forks, resulting in replicative stress. By siRNA screening, we identified kinases involved in the accumulation of phosphohistone 2AX (γH2AX) upon UV irradiation-induced replication stress. Surprisingly, the strongest reduction of phosphohistone 2AX followed knockdown of the MAP kinase-activated protein kinase 2 (MK2), a kinase currently implicated in p38 stress signaling and G2 arrest. Depletion or inhibition of MK2 also protected cells from DNA damage-induced cell death, and mice deficient for MK2 displayed decreased apoptosis in the skin upon UV irradiation. Moreover, MK2 activity was required for damage response, accumulation of ssDNA, and decreased survival when cells were treated with the nucleoside analogue gemcitabine or when the checkpoint kinase Chk1 was antagonized. By using DNA fiber assays, we found that MK2 inhibition or knockdown rescued DNA replication impaired by gemcitabine or by Chk1 inhibition. This rescue strictly depended on translesion DNA polymerases. In conclusion, instead of being an unavoidable consequence of DNA damage, alterations of replication speed and origin firing depend on MK2-mediated signaling.

KW - Animals

KW - Antimetabolites, Antineoplastic

KW - Cell Line, Tumor

KW - DNA Damage

KW - DNA Replication

KW - DNA, Single-Stranded

KW - Deoxycytidine

KW - G2 Phase Cell Cycle Checkpoints

KW - Gene Knockdown Techniques

KW - Histones

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - MAP Kinase Signaling System

KW - Mice

KW - Mice, Knockout

KW - Protein Kinases

KW - Protein-Serine-Threonine Kinases

KW - Ultraviolet Rays

KW - p38 Mitogen-Activated Protein Kinases

U2 - 10.1073/pnas.1304355110

DO - 10.1073/pnas.1304355110

M3 - Journal article

C2 - 24082115

SN - 0027-8424

VL - 110

SP - 16856

EP - 16861

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 42

ER -