RNF8 and RNF168 but not HERC2 are required for DNA damage-induced ubiquitylation in chicken DT40 cells

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Vibe Hallundbæk Østergaard, Constanze Pentzold, Rune Troelsgaard Pedersen, Silviu Iosif, Arno Alpi, Simon Bekker-Jensen, Niels Mailand, Michael Lisby

The ubiquitylation cascade plays an important role in the recruitment of repair factors at DNA double-strand breaks. The involvement of a growing number of ubiquitin E3 ligases adds to the complexity of the DNA damage-induced ubiquitin signaling. Here we use the genetically tractable avian cell line DT40 to investigate the role of HERC2, RNF8 and RNF168 in the DNA damage-induced ubiquitylation pathway. We show that formation of ubiquitin foci as well as cell survival after DNA damage depends on both RNF8 and RNF168. However, we find that RNF8 and RNF168 knockout cell lines respond differently to treatment with camptothecin indicating that they do not function in a strictly linear manner. Surprisingly, we show that HERC2 is required neither for survival nor for ubiquitin foci formation after DNA damage in DT40. Moreover, the E3 ubiquitin ligase activity of HERC2 is not redundant to that of RNF8 or RNF168.
OriginalsprogEngelsk
TidsskriftD N A Repair
Vol/bind11
Udgave nummer11
Sider (fra-til)892–905
Antal sider14
ISSN1568-7864
DOI
StatusUdgivet - 2012

ID: 41804581