The C-terminal domain (CTD) of human DNA glycosylase NEIL1 is required for forming BERosome repair complex with DNA replication proteins at the replicating genome

Dominant negative function of the CTD

Pavana M. Hegde, Arijit Dutta, Shiladitya Sengupta, Joy Mitra, Sanjay Adhikari, Alan E. Tomkinson, Guo Min Li, Istvan Boldogh, Tapas Hazra, Sankar Mitra, Muralidhar L. Hegde

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The human DNA glycosylase NEIL1 was recently demonstrated to initiate prereplicative base excision repair (BER) of oxidized bases in the replicating genome, thus preventing mutagenic replication. A significant fraction of NEIL1 in cells is present in large cellular complexes containing DNA replication and other repair proteins, as shown by gel filtration. However, how the interaction of NEIL1 affects its recruitment to the replication site for prereplicative repair was not investigated. Here, we show that NEIL1 binarily interacts with the proliferating cell nuclear antigen clamp loader replication factor C, DNA polymerase δ, and DNA ligase I in the absence of DNA via its nonconserved C-terminal domain (CTD); replication factor C interaction results in ∼8-fold stimulation of NEIL1 activity. Disruption of NEIL1 interactions within the BERosome complex, as observed for a NEIL1 deletion mutant (N311) lacking the CTD, not only inhibits complete BER in vitro but also prevents its chromatin association and reduced recruitment at replication foci in S phase cells. This suggests that the interaction of NEIL1 with replication and other BER proteins is required for efficient repair of the replicating genome. Consistently, the CTD polypeptide acts as a dominant negative inhibitor during in vitro repair, and its ectopic expression sensitizes human cells to reactive oxygen species. We conclude that multiple interactions among BER proteins lead to large complexes, which are critical for efficient BER in mammalian cells, and the CTD interaction could be targeted for enhancing drug/radiation sensitivity of tumor cells.

Original languageEnglish (US)
Pages (from-to)20919-20933
Number of pages15
JournalJournal of Biological Chemistry
Volume290
Issue number34
DOIs
StatePublished - Aug 21 2015

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DNA Glycosylases
DNA Replication
DNA Repair
Repair
Genes
Genome
Replication Protein C
DNA
Proteins
Cells
Radiation Tolerance
Proliferating Cell Nuclear Antigen
DNA-Directed DNA Polymerase
S Phase
Chromatin
Gel Chromatography
Reactive Oxygen Species
DNA Ligases
Loaders
Peptides

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

The C-terminal domain (CTD) of human DNA glycosylase NEIL1 is required for forming BERosome repair complex with DNA replication proteins at the replicating genome : Dominant negative function of the CTD. / Hegde, Pavana M.; Dutta, Arijit; Sengupta, Shiladitya; Mitra, Joy; Adhikari, Sanjay; Tomkinson, Alan E.; Li, Guo Min; Boldogh, Istvan; Hazra, Tapas; Mitra, Sankar; Hegde, Muralidhar L.

In: Journal of Biological Chemistry, Vol. 290, No. 34, 21.08.2015, p. 20919-20933.

Research output: Contribution to journalArticle

Hegde, Pavana M. ; Dutta, Arijit ; Sengupta, Shiladitya ; Mitra, Joy ; Adhikari, Sanjay ; Tomkinson, Alan E. ; Li, Guo Min ; Boldogh, Istvan ; Hazra, Tapas ; Mitra, Sankar ; Hegde, Muralidhar L. / The C-terminal domain (CTD) of human DNA glycosylase NEIL1 is required for forming BERosome repair complex with DNA replication proteins at the replicating genome : Dominant negative function of the CTD. In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 34. pp. 20919-20933.
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AU - Dutta, Arijit

AU - Sengupta, Shiladitya

AU - Mitra, Joy

AU - Adhikari, Sanjay

AU - Tomkinson, Alan E.

AU - Li, Guo Min

AU - Boldogh, Istvan

AU - Hazra, Tapas

AU - Mitra, Sankar

AU - Hegde, Muralidhar L.

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