TY - JOUR
T1 - Histone H2A variants alpha1-extension helix directs RNF168-mediated ubiquitination
AU - Kelliher, Jessica L.
AU - West, Kirk L.
AU - Gong, Qingguo
AU - Leung, Justin W.C.
N1 - Funding Information:
We thank Drs. Fen Xia (UAMS), Tanya Paull (UT Austin), Ilya Finkelstein (UT Austin), and John Pehrson (U Penn) for the insightful discussion. We would also like to thank Alicia Byrd (UAMS), Drs. Uma Muthurajan and Hataichanok Scherman (Colorado State University Histone Source) for the technical supports on nucleosome reconstitution, the UAMS OSPAN Science Communication (SciCom) group for editing the manuscript. This work was supported by grants from the NIH (K22CA204354) and Arkansas Breast Cancer Research Program (AWD00053730) to J.W.L. and in part by start-up funds from the University of Arkansas for Medical Sciences and NIH (P20GM121293). The National Natural Science Foundation of China (Grant 31770805) to Q.G.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Histone ubiquitination plays an important role in the DNA damage response (DDR) pathway. RNF168 catalyzes H2A and H2AX ubiquitination on lysine 13/15 (K13/K15) upon DNA damage and promotes the accrual of downstream repair factors at damaged chromatin. Here, we report that RNF168 ubiquitinates the non-canonical H2A variants H2AZ and macroH2A1/2 at the divergent N-terminal tail lysine residue. In addition to their evolutionarily conserved nucleosome acidic patch, we identify the positively charged alpha1-extension helix as essential for RNF168-mediated ubiquitination of H2A variants. Moreover, mutation of the RNF168 UMI (UIM- and MIU-related UBD) hydrophilic acidic residues abolishes RNF168-mediated ubiquitination as well as 53BP1 and BRCA1 ionizing radiation-induced foci formation. Our results reveal a juxtaposed bipartite electrostatic interaction utilized by the nucleosome to direct RNF168 orientation towards the target lysine residues in proximity to the H2A alpha1-extension helix, which plays an important role in the DDR pathway.
AB - Histone ubiquitination plays an important role in the DNA damage response (DDR) pathway. RNF168 catalyzes H2A and H2AX ubiquitination on lysine 13/15 (K13/K15) upon DNA damage and promotes the accrual of downstream repair factors at damaged chromatin. Here, we report that RNF168 ubiquitinates the non-canonical H2A variants H2AZ and macroH2A1/2 at the divergent N-terminal tail lysine residue. In addition to their evolutionarily conserved nucleosome acidic patch, we identify the positively charged alpha1-extension helix as essential for RNF168-mediated ubiquitination of H2A variants. Moreover, mutation of the RNF168 UMI (UIM- and MIU-related UBD) hydrophilic acidic residues abolishes RNF168-mediated ubiquitination as well as 53BP1 and BRCA1 ionizing radiation-induced foci formation. Our results reveal a juxtaposed bipartite electrostatic interaction utilized by the nucleosome to direct RNF168 orientation towards the target lysine residues in proximity to the H2A alpha1-extension helix, which plays an important role in the DDR pathway.
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U2 - 10.1038/s41467-020-16307-4
DO - 10.1038/s41467-020-16307-4
M3 - Article
C2 - 32424115
AN - SCOPUS:85084787906
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 2462
ER -