The role of HERC2 and RNF8 ubiquitin E3 ligases in the promotion of translesion DNA synthesis in the chicken DT40 cell line

[Unknown] Mohiuddin, Shunsuke Kobayashi, Islam Shamima Keka, Guillaume Guilbaud, Julian Sale, Takeo Narita, H Ismail Abdel-Aziz, Xin Wang, Saki Ogawa, Hiroyuki Sasanuma, Roland Chiu, Vibe H Oestergaard, Michael Lisby, Shunichi Takeda

Research output: Contribution to journalArticleAcademicpeer-review

12 Citations (Scopus)
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Abstract

The replicative DNA polymerases are generally blocked by template DNA damage. The resulting replication arrest can be released by one of two post-replication repair (PRR) pathways, translesion DNA synthesis (TLS) and template switching by homologous recombination (HR). The HERC2 ubiquitin ligase plays a role in homologous recombination by facilitating the assembly of the Ubc13 ubiquitin-conjugating enzyme with the RNF8 ubiquitin ligase. To explore the role of HERC2 and RNF8 in PRR, we examined immunoglobulin diversification in chicken DT40 cells deficient in HERC2 and RNF8. Unexpectedly, the HERC2(-/-) and RNF8(-/-) cells and HERC2(-/-)/RNF8(-/-) double mutant cells exhibit a significant reduction in the rate of immunoglobulin (Ig) hypermutation, compared to wild-type cells. Further, the HERC2(-/-) and RNF8(-/-) mutants exhibit defective maintenance of replication fork progression immediately after exposure to UV while retaining proficient post-replicative gap filling. These mutants are both proficient in mono-ubiquitination of PCNA. Taken together, these results suggest that HERC2 and RNF8 promote TLS past abasic sites and UV-lesions at or very close to stalled replication forks.

Original languageEnglish
Pages (from-to)67-76
Number of pages10
JournalDna repair
Volume40
DOIs
Publication statusPublished - Apr-2016

Keywords

  • Animals
  • Chickens
  • DNA Damage
  • DNA Replication
  • Guanine Nucleotide Exchange Factors
  • HEK293 Cells
  • Humans
  • Mutation
  • Ubiquitin-Protein Ligases
  • Ubiquitination
  • Ultraviolet Rays
  • Journal Article
  • Research Support, Non-U.S. Gov't

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