@article{f4fb20dc985e4c7a8e01b663271bfe28,
title = "Ubiquitin Phosphorylation at Thr12 Modulates the DNA Damage Response",
abstract = "The ubiquitin system regulates the DNA damage response (DDR) by modifying histone H2A at Lys15 (H2AK15ub) and triggering downstream signaling events. Here, we find that phosphorylation of ubiquitin at Thr12 (pUbT12) controls the DDR by inhibiting the function of 53BP1, a key factor for DNA double-strand break repair by non-homologous end joining (NHEJ). Detectable as a chromatin modification on H2AK15ub, pUbT12 accumulates in nuclear foci and is increased upon DNA damage. Mutating Thr12 prevents the removal of ubiquitin from H2AK15ub by USP51 deubiquitinating enzyme, leading to a pronounced accumulation of ubiquitinated chromatin. Chromatin modified by pUbT12 is inaccessible to 53BP1 but permissive to the homologous recombination (HR) proteins RNF169, RAD51, and the BRCA1/BARD1 complex. Phosphorylation of ubiquitin at Thr12 in the chromatin context is a new histone mark, H2AK15pUbT12, that regulates the DDR by hampering the activity of 53BP1 at damaged chromosomes.",
keywords = "53BP1, BRCA1/BARD1, DDR, DNA damage response, DNA repair, H2AK15pUbT12, RAD51, RNF168, RNF169, RNF8, USP51, chromatin ubiquitination, genome stability, histone mark H2AK15ub, pUbT12, phospho-ubiquitin Thr12, ubiquitin phosphorylation",
author = "Franziska Walser and Mulder, {Monique P.C.} and Beno{\^i}t Bragantini and Sibylle Burger and Tatiana Gubser and Marco Gatti and Botuyan, {Maria Victoria} and Alessandra Villa and Matthias Altmeyer and Dario Neri and Huib Ovaa and Georges Mer and Lorenza Penengo",
note = "Funding Information: We thank Michael Walser, Beat Sch?fer, Jeremy Stark, Richard Baer, Alessandro Sartori, Michael Huen, and David Komander for sharing useful reagents; Qi Hu for molecular modeling; Eliana Bianco and Stefano Ferrari for technical support; the Center for Microscopy and Image Analysis of the University of Zurich for technical assistance with imaging; and Richard Chahwan for discussion. This work was supported by the National Institutes of Health (NIH; grants CA132878 and GM116829) to G.M. the Netherlands Foundation for Scientific Research (VICI; grant 724.013.002) to H.O. the Swiss National Science Foundation (SNSF; grants 310030_184966 and 31003A_166370), the Novartis Foundation (grant 17A039) and the Krebsliga (grant KFS-4577-08-2018) to L.P. F.W. performed most of the experiments. M.P.C.M. and H.O. synthetized phosphorylated ubiquitin. B.B. M.V.B. and G.M. produced ubiquitinated nucleosomes and performed pull-downs. S.B. contributed to immunofluorescence studies and olaparib sensitivity assays. T.G. generated the ubiquitin replacement system. A.V. and D.N. provided expertise and reagents for the phage display selection. M.G. and M.A. contributed to quantitative imaging. L.P. conceived the project, designed experiments, and wrote the manuscript, supported by F.W. H.O. is a shareholder of UbiQ B.V. Funding Information: We thank Michael Walser, Beat Sch{\"a}fer, Jeremy Stark, Richard Baer, Alessandro Sartori, Michael Huen, and David Komander for sharing useful reagents; Qi Hu for molecular modeling; Eliana Bianco and Stefano Ferrari for technical support; the Center for Microscopy and Image Analysis of the University of Zurich for technical assistance with imaging; and Richard Chahwan for discussion. This work was supported by the National Institutes of Health (NIH; grants CA132878 and GM116829) to G.M., the Netherlands Foundation for Scientific Research (VICI; grant 724.013.002) to H.O., the Swiss National Science Foundation (SNSF; grants 310030_184966 and 31003A_166370) , the Novartis Foundation (grant 17A039) and the Krebsliga (grant KFS-4577-08-2018) to L.P. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = nov,
day = "5",
doi = "10.1016/j.molcel.2020.09.017",
language = "English (US)",
volume = "80",
pages = "423--436.e9",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "3",
}