4AYC

RNF8 RING domain structure

Summary for 4AYC

• Entry DOI: 10.2210/pdb4ayc/pdb
• Related: 2CSW
• Descriptor: E3 UBIQUITIN-PROTEIN LIGASE RNF8, SULFATE ION, N,N-BIS(3-D-GLUCONAMIDOPROPYL)DEOXYCHOLAMIDE, ... (8 entities in total)
• Functional Keywords: ligase, dna damage, k63 chains
• Biological source: HOMO SAPIENS (HUMAN); More
• Cellular location: Nucleus: O76064 O76064
• Total number of polymer chains: 2
• Total formula weight: 34261.59

Authors

Mattiroli, F.,Vissers, J.H.A.,Van Dijk, W.J.,Ikpa, P.,Citterio, E.,Vermeulen, W.,Marteijn, J.A.,Sixma, T.K. (deposition date: 2012-06-20, release date: 2012-09-26, Last modification date: 2024-11-13)

Primary citation

Mattiroli, F.,Vissers, J.H.A.,Van Dijk, W.J.,Ikpa, P.,Citterio, E.,Vermeulen, W.,Marteijn, J.A.,Sixma, T.K.
Rnf168 Ubiquitinates K13-15 on H2A/H2Ax to Drive DNA Damage Signaling
Cell(Cambridge,Mass.), 150:1182-, 2012

PubMed Abstract: Ubiquitin-dependent signaling during the DNA damage response (DDR) to double-strand breaks (DSBs) is initiated by two E3 ligases, RNF8 and RNF168, targeting histone H2A and H2AX. RNF8 is the first ligase recruited to the damage site, and RNF168 follows RNF8-dependent ubiquitination. This suggests that RNF8 initiates H2A/H2AX ubiquitination with K63-linked ubiquitin chains and RNF168 extends them. Here, we show that RNF8 is inactive toward nucleosomal H2A, whereas RNF168 catalyzes the monoubiquitination of the histones specifically on K13-15. Structure-based mutagenesis of RNF8 and RNF168 RING domains shows that a charged residue determines whether nucleosomal proteins are recognized. We find that K63 ubiquitin chains are conjugated to RNF168-dependent H2A/H2AX monoubiquitination at K13-15 and not on K118-119. Using a mutant of RNF168 unable to target histones but still catalyzing ubiquitin chains at DSBs, we show that ubiquitin chains per se are insufficient for signaling, but RNF168 target ubiquitination is required for DDR.

PubMed: 22980979
DOI: 10.1016/J.CELL.2012.08.005

Experimental method

X-RAY DIFFRACTION (1.9 Å)