Acetylation of oxidized base repair-initiating NEIL1 DNA glycosylase required for chromatin-bound repair complex formation in the human genome increases cellular resistance to oxidative stress

Shiladitya Sengupta, Chunying Yang, Muralidhar L. Hegde, Pavana M. Hegde, Joy Mitra, Arvind Pandey, Arijit Dutta, Abdul Tayyeb Datarwala, Kishor K Bhakat, Sankar Mitra

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Posttranslational modifications of DNA repair proteins have been linked to their function. However, it is not clear if posttranslational acetylation affects subcellular localization of these enzymes. Here, we show that the human DNA glycosylase NEIL1, which is involved in repair of both endo- and exogenously generated oxidized bases via the base excision repair (BER) pathway, is acetylated by histone acetyltransferase p300. Acetylation occurs predominantly at Lys residues 296, 297 and 298 located in NEIL1’s disordered C-terminal domain. NEIL1 mutant having the substitution of Lys 296–298 with neutral Ala loses nuclear localization, whereas Lys > Arg substitution (in 3KR mutant) at the same sites does not affect NEIL1’s nuclear localization or chromatin binding, presumably due to retention of the positive charge. Although non-acetylated NEIL1 can bind to chromatin, acetylated NEIL1 is exclusively chromatin-bound. NEIL1 acetylation while dispensable for its glycosylase activity enhances it due to increased product release. The acetylation-defective 3KR mutant forms less stable complexes with various chromatin proteins, including histone chaperones and BER/single-strand break repair partners, than the wild-type (WT) NEIL1. We also showed that the repair complex with WT NEIL1 has significantly higher BER activity than the 3KR mutant complex. This is consistent with reduced resistance of non-acetylable mutant NEIL1 expressing cells to oxidative stress relative to cells expressing the acetylable WT enzyme. We thus conclude that the major role of acetylable Lys residues in NEIL1 is to stabilize the formation of chromatin-bound repair complexes which protect cells from oxidative stress.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalDNA Repair
Volume66-67
DOIs
StatePublished - Jun 1 2018

Fingerprint

DNA Glycosylases
Acetylation
Oxidative stress
Human Genome
Chromatin
Oxidative Stress
Repair
Genes
DNA Repair
Histone Chaperones
Histone Acetyltransferases
Enzymes
Post Translational Protein Processing
Substitution reactions
Proteins
Cells
DNA

Keywords

  • Acetylation
  • Base excision repair (BER)
  • Chromatin
  • NEIL1
  • Oxidative stress
  • Repair complex

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Acetylation of oxidized base repair-initiating NEIL1 DNA glycosylase required for chromatin-bound repair complex formation in the human genome increases cellular resistance to oxidative stress. / Sengupta, Shiladitya; Yang, Chunying; Hegde, Muralidhar L.; Hegde, Pavana M.; Mitra, Joy; Pandey, Arvind; Dutta, Arijit; Datarwala, Abdul Tayyeb; Bhakat, Kishor K; Mitra, Sankar.

In: DNA Repair, Vol. 66-67, 01.06.2018, p. 1-10.

Research output: Contribution to journalArticle

Sengupta, Shiladitya ; Yang, Chunying ; Hegde, Muralidhar L. ; Hegde, Pavana M. ; Mitra, Joy ; Pandey, Arvind ; Dutta, Arijit ; Datarwala, Abdul Tayyeb ; Bhakat, Kishor K ; Mitra, Sankar. / Acetylation of oxidized base repair-initiating NEIL1 DNA glycosylase required for chromatin-bound repair complex formation in the human genome increases cellular resistance to oxidative stress. In: DNA Repair. 2018 ; Vol. 66-67. pp. 1-10.
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abstract = "Posttranslational modifications of DNA repair proteins have been linked to their function. However, it is not clear if posttranslational acetylation affects subcellular localization of these enzymes. Here, we show that the human DNA glycosylase NEIL1, which is involved in repair of both endo- and exogenously generated oxidized bases via the base excision repair (BER) pathway, is acetylated by histone acetyltransferase p300. Acetylation occurs predominantly at Lys residues 296, 297 and 298 located in NEIL1’s disordered C-terminal domain. NEIL1 mutant having the substitution of Lys 296–298 with neutral Ala loses nuclear localization, whereas Lys > Arg substitution (in 3KR mutant) at the same sites does not affect NEIL1’s nuclear localization or chromatin binding, presumably due to retention of the positive charge. Although non-acetylated NEIL1 can bind to chromatin, acetylated NEIL1 is exclusively chromatin-bound. NEIL1 acetylation while dispensable for its glycosylase activity enhances it due to increased product release. The acetylation-defective 3KR mutant forms less stable complexes with various chromatin proteins, including histone chaperones and BER/single-strand break repair partners, than the wild-type (WT) NEIL1. We also showed that the repair complex with WT NEIL1 has significantly higher BER activity than the 3KR mutant complex. This is consistent with reduced resistance of non-acetylable mutant NEIL1 expressing cells to oxidative stress relative to cells expressing the acetylable WT enzyme. We thus conclude that the major role of acetylable Lys residues in NEIL1 is to stabilize the formation of chromatin-bound repair complexes which protect cells from oxidative stress.",
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AU - Sengupta, Shiladitya

AU - Yang, Chunying

AU - Hegde, Muralidhar L.

AU - Hegde, Pavana M.

AU - Mitra, Joy

AU - Pandey, Arvind

AU - Dutta, Arijit

AU - Datarwala, Abdul Tayyeb

AU - Bhakat, Kishor K

AU - Mitra, Sankar

PY - 2018/6/1

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AB - Posttranslational modifications of DNA repair proteins have been linked to their function. However, it is not clear if posttranslational acetylation affects subcellular localization of these enzymes. Here, we show that the human DNA glycosylase NEIL1, which is involved in repair of both endo- and exogenously generated oxidized bases via the base excision repair (BER) pathway, is acetylated by histone acetyltransferase p300. Acetylation occurs predominantly at Lys residues 296, 297 and 298 located in NEIL1’s disordered C-terminal domain. NEIL1 mutant having the substitution of Lys 296–298 with neutral Ala loses nuclear localization, whereas Lys > Arg substitution (in 3KR mutant) at the same sites does not affect NEIL1’s nuclear localization or chromatin binding, presumably due to retention of the positive charge. Although non-acetylated NEIL1 can bind to chromatin, acetylated NEIL1 is exclusively chromatin-bound. NEIL1 acetylation while dispensable for its glycosylase activity enhances it due to increased product release. The acetylation-defective 3KR mutant forms less stable complexes with various chromatin proteins, including histone chaperones and BER/single-strand break repair partners, than the wild-type (WT) NEIL1. We also showed that the repair complex with WT NEIL1 has significantly higher BER activity than the 3KR mutant complex. This is consistent with reduced resistance of non-acetylable mutant NEIL1 expressing cells to oxidative stress relative to cells expressing the acetylable WT enzyme. We thus conclude that the major role of acetylable Lys residues in NEIL1 is to stabilize the formation of chromatin-bound repair complexes which protect cells from oxidative stress.

KW - Acetylation

KW - Base excision repair (BER)

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KW - Oxidative stress

KW - Repair complex

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