TY - JOUR
T1 - DNA damage-inducible SUMOylation of HERC2 promotes RNF8 binding via a novel SUMO-binding Zinc finger
AU - Danielsen, Jannie Michaela Rendtlew
AU - Povlsen, Lou Klitgaard
AU - Villumsen, Bine Hare
AU - Streicher, Werner
AU - Nilsson, Jakob
AU - Wikström, Mats
AU - Bekker-Jensen, Simon
AU - Mailand, Niels
PY - 2012/4
Y1 - 2012/4
N2 - Nonproteolytic ubiquitylation of chromatin surrounding deoxyribonucleic acid (DNA) double-strand breaks (DSBs) by the RNF8/RNF168/HERC2 ubiquitin ligases facilitates restoration of genome integrity by licensing chromatin to concentrate genome caretaker proteins near the lesions. In parallel, SUMOylation of so-far elusive upstream DSB regulators is also required for execution of this ubiquitin-dependent chromatin response. We show that HERC2 and RNF168 are novel DNA damage-dependent SUMOylation targets in human cells. In response to DSBs, both HERC2 and RNF168 were specifically modified with SUMO1 at DSB sites in a manner dependent on the SUMO E3 ligase PIAS4. SUMOylation of HERC2 was required for its DSB-induced association with RNF8 and for stabilizing the RNF8-Ubc13 complex. We also demonstrate that the ZZ Zinc finger in HERC2 defined a novel SUMO-specific binding module, which together with its concomitant SUMOylation and T4827 phosphorylation promoted binding to RNF8. Our findings provide novel insight into the regulatory complexity of how ubiquitylation and SUMOylation cooperate to orchestrate protein interactions with DSB repair foci.
AB - Nonproteolytic ubiquitylation of chromatin surrounding deoxyribonucleic acid (DNA) double-strand breaks (DSBs) by the RNF8/RNF168/HERC2 ubiquitin ligases facilitates restoration of genome integrity by licensing chromatin to concentrate genome caretaker proteins near the lesions. In parallel, SUMOylation of so-far elusive upstream DSB regulators is also required for execution of this ubiquitin-dependent chromatin response. We show that HERC2 and RNF168 are novel DNA damage-dependent SUMOylation targets in human cells. In response to DSBs, both HERC2 and RNF168 were specifically modified with SUMO1 at DSB sites in a manner dependent on the SUMO E3 ligase PIAS4. SUMOylation of HERC2 was required for its DSB-induced association with RNF8 and for stabilizing the RNF8-Ubc13 complex. We also demonstrate that the ZZ Zinc finger in HERC2 defined a novel SUMO-specific binding module, which together with its concomitant SUMOylation and T4827 phosphorylation promoted binding to RNF8. Our findings provide novel insight into the regulatory complexity of how ubiquitylation and SUMOylation cooperate to orchestrate protein interactions with DSB repair foci.
KW - Cell Line
KW - Chromatin
KW - DNA Breaks, Double-Stranded
KW - DNA Damage
KW - DNA Repair
KW - DNA-Binding Proteins
KW - Guanine Nucleotide Exchange Factors
KW - HEK293 Cells
KW - Humans
KW - Protein Inhibitors of Activated STAT
KW - RNA Interference
KW - RNA, Small Interfering
KW - SUMO-1 Protein
KW - Sumoylation
KW - Ubiquitin-Conjugating Enzymes
KW - Ubiquitin-Protein Ligases
KW - Zinc Fingers
U2 - 10.1083/jcb.201106152
DO - 10.1083/jcb.201106152
M3 - Journal article
C2 - 22508508
SN - 0021-9525
VL - 197
SP - 179
EP - 187
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 2
ER -