RNF8 ubiquitylates histones at DNA double-strand breaks and promotes assembly of repair proteins

TY - JOUR

T1 - RNF8 ubiquitylates histones at DNA double-strand breaks and promotes assembly of repair proteins

AU - Mailand, Niels

AU - Bekker-Jensen, Simon

AU - Faustrup, Helene

AU - Melander, Fredrik

AU - Bartek, Jiri

AU - Lukas, Claudia

AU - Lukas, Jiri

PY - 2007

Y1 - 2007

N2 - Accumulation of repair proteins on damaged chromosomes is required to restore genomic integrity. However, the mechanisms of protein retention at the most destructive chromosomal lesions, the DNA double-strand breaks (DSBs), are poorly understood. We show that RNF8, a RING-finger ubiquitin ligase, rapidly assembles at DSBs via interaction of its FHA domain with the phosphorylated adaptor protein MDC1. This is accompanied by an increase in DSB-associated ubiquitylations and followed by accumulation of 53BP1 and BRCA1 repair proteins. Knockdown of RNF8 or disruption of its FHA or RING domains impaired DSB-associated ubiquitylation and inhibited retention of 53BP1 and BRCA1 at the DSB sites. In addition, we show that RNF8 can ubiquitylate histone H2A and H2AX, and that its depletion sensitizes cells to ionizing radiation. These data suggest that MDC1-mediated and RNF8-executed histone ubiquitylation protects genome integrity by licensing the DSB-flanking chromatin to concentrate repair factors near the DNA lesions.

AB - Accumulation of repair proteins on damaged chromosomes is required to restore genomic integrity. However, the mechanisms of protein retention at the most destructive chromosomal lesions, the DNA double-strand breaks (DSBs), are poorly understood. We show that RNF8, a RING-finger ubiquitin ligase, rapidly assembles at DSBs via interaction of its FHA domain with the phosphorylated adaptor protein MDC1. This is accompanied by an increase in DSB-associated ubiquitylations and followed by accumulation of 53BP1 and BRCA1 repair proteins. Knockdown of RNF8 or disruption of its FHA or RING domains impaired DSB-associated ubiquitylation and inhibited retention of 53BP1 and BRCA1 at the DSB sites. In addition, we show that RNF8 can ubiquitylate histone H2A and H2AX, and that its depletion sensitizes cells to ionizing radiation. These data suggest that MDC1-mediated and RNF8-executed histone ubiquitylation protects genome integrity by licensing the DSB-flanking chromatin to concentrate repair factors near the DNA lesions.

KW - BRCA1 Protein

KW - Binding Sites

KW - Cell Survival

KW - Chromatin

KW - DNA Breaks, Double-Stranded

KW - DNA Repair Enzymes

KW - DNA-Binding Proteins

KW - Histones

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - Models, Biological

KW - Nuclear Proteins

KW - Phosphorylation

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Trans-Activators

KW - Tumor Cells, Cultured

KW - Ubiquitination

U2 - 10.1016/j.cell.2007.09.040

DO - 10.1016/j.cell.2007.09.040

M3 - Journal article

C2 - 18001824

SN - 0092-8674

VL - 131

SP - 887

EP - 900

JO - Cell

JF - Cell

IS - 5

ER -