Preferential repair of oxidized base damage in the transcribed genes of mammalian cells

Dibyendu Banerjee, Santi M. Mandal, Aditi Das, Muralidhar L. Hegde, Soumita Das, Kishor K. Bhakat, Istvan Boldogh, Partha Sarkar, Sankar Mitra, Tapas Hazra

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Preferential repair of bulky DNA adducts from the transcribed genes via nucleotide excision repair is well characterized in mammalian cells. However, definitive evidence is lacking for similar repair of oxidized bases, the major endogenous DNA lesions. Here we show that the oxidized base-specific human DNA glycosylase NEIL2 associates with RNA polymerase II and the transcriptional regulator heterogeneous nuclear ribonucleoprotein-U (hnRNP-U), both in vitro and in cells. NEIL2 immunocomplexes from cell extracts preferentially repaired the mutagenic cytosine oxidation product 5-hydroxyuracil in the transcribed strand. In a reconstituted system, we also observed NEIL2-initiated transcription- dependent base excision repair of 5-hydroxyuracil in the transcribed strand, with hnRNP-U playing a critical role. Chromatin immunoprecipitation/ reimmunoprecipitation studies showed association of NEIL2, RNA polymerase II, and hnRNP-U on transcribed but not on transcriptionally silent genes. Furthermore, NEIL2-depleted cells accumulated more DNA damage in active than in silent genes. These results strongly support the preferential role of NEIL2 in repairing oxidized bases in the transcribed genes of mammalian cells.

Original languageEnglish (US)
Pages (from-to)6006-6016
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number8
DOIs
StatePublished - Feb 25 2011

Fingerprint

Heterogeneous-Nuclear Ribonucleoprotein U
Repair
Genes
Cells
RNA Polymerase II
DNA Repair
DNA Glycosylases
DNA Adducts
Chromatin Immunoprecipitation
Cytosine
DNA
Transcription
Cell Extracts
DNA Damage
Chromatin
Nucleotides
Association reactions
Oxidation
5-hydroxyuracil

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Preferential repair of oxidized base damage in the transcribed genes of mammalian cells. / Banerjee, Dibyendu; Mandal, Santi M.; Das, Aditi; Hegde, Muralidhar L.; Das, Soumita; Bhakat, Kishor K.; Boldogh, Istvan; Sarkar, Partha; Mitra, Sankar; Hazra, Tapas.

In: Journal of Biological Chemistry, Vol. 286, No. 8, 25.02.2011, p. 6006-6016.

Research output: Contribution to journalArticle

Banerjee, Dibyendu ; Mandal, Santi M. ; Das, Aditi ; Hegde, Muralidhar L. ; Das, Soumita ; Bhakat, Kishor K. ; Boldogh, Istvan ; Sarkar, Partha ; Mitra, Sankar ; Hazra, Tapas. / Preferential repair of oxidized base damage in the transcribed genes of mammalian cells. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 8. pp. 6006-6016.
@article{76771ee91aa345c2b866a04a94326341,
title = "Preferential repair of oxidized base damage in the transcribed genes of mammalian cells",
abstract = "Preferential repair of bulky DNA adducts from the transcribed genes via nucleotide excision repair is well characterized in mammalian cells. However, definitive evidence is lacking for similar repair of oxidized bases, the major endogenous DNA lesions. Here we show that the oxidized base-specific human DNA glycosylase NEIL2 associates with RNA polymerase II and the transcriptional regulator heterogeneous nuclear ribonucleoprotein-U (hnRNP-U), both in vitro and in cells. NEIL2 immunocomplexes from cell extracts preferentially repaired the mutagenic cytosine oxidation product 5-hydroxyuracil in the transcribed strand. In a reconstituted system, we also observed NEIL2-initiated transcription- dependent base excision repair of 5-hydroxyuracil in the transcribed strand, with hnRNP-U playing a critical role. Chromatin immunoprecipitation/ reimmunoprecipitation studies showed association of NEIL2, RNA polymerase II, and hnRNP-U on transcribed but not on transcriptionally silent genes. Furthermore, NEIL2-depleted cells accumulated more DNA damage in active than in silent genes. These results strongly support the preferential role of NEIL2 in repairing oxidized bases in the transcribed genes of mammalian cells.",
author = "Dibyendu Banerjee and Mandal, {Santi M.} and Aditi Das and Hegde, {Muralidhar L.} and Soumita Das and Bhakat, {Kishor K.} and Istvan Boldogh and Partha Sarkar and Sankar Mitra and Tapas Hazra",
year = "2011",
month = "2",
day = "25",
doi = "10.1074/jbc.M110.198796",
language = "English (US)",
volume = "286",
pages = "6006--6016",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "8",

}

TY - JOUR

T1 - Preferential repair of oxidized base damage in the transcribed genes of mammalian cells

AU - Banerjee, Dibyendu

AU - Mandal, Santi M.

AU - Das, Aditi

AU - Hegde, Muralidhar L.

AU - Das, Soumita

AU - Bhakat, Kishor K.

AU - Boldogh, Istvan

AU - Sarkar, Partha

AU - Mitra, Sankar

AU - Hazra, Tapas

PY - 2011/2/25

Y1 - 2011/2/25

N2 - Preferential repair of bulky DNA adducts from the transcribed genes via nucleotide excision repair is well characterized in mammalian cells. However, definitive evidence is lacking for similar repair of oxidized bases, the major endogenous DNA lesions. Here we show that the oxidized base-specific human DNA glycosylase NEIL2 associates with RNA polymerase II and the transcriptional regulator heterogeneous nuclear ribonucleoprotein-U (hnRNP-U), both in vitro and in cells. NEIL2 immunocomplexes from cell extracts preferentially repaired the mutagenic cytosine oxidation product 5-hydroxyuracil in the transcribed strand. In a reconstituted system, we also observed NEIL2-initiated transcription- dependent base excision repair of 5-hydroxyuracil in the transcribed strand, with hnRNP-U playing a critical role. Chromatin immunoprecipitation/ reimmunoprecipitation studies showed association of NEIL2, RNA polymerase II, and hnRNP-U on transcribed but not on transcriptionally silent genes. Furthermore, NEIL2-depleted cells accumulated more DNA damage in active than in silent genes. These results strongly support the preferential role of NEIL2 in repairing oxidized bases in the transcribed genes of mammalian cells.

AB - Preferential repair of bulky DNA adducts from the transcribed genes via nucleotide excision repair is well characterized in mammalian cells. However, definitive evidence is lacking for similar repair of oxidized bases, the major endogenous DNA lesions. Here we show that the oxidized base-specific human DNA glycosylase NEIL2 associates with RNA polymerase II and the transcriptional regulator heterogeneous nuclear ribonucleoprotein-U (hnRNP-U), both in vitro and in cells. NEIL2 immunocomplexes from cell extracts preferentially repaired the mutagenic cytosine oxidation product 5-hydroxyuracil in the transcribed strand. In a reconstituted system, we also observed NEIL2-initiated transcription- dependent base excision repair of 5-hydroxyuracil in the transcribed strand, with hnRNP-U playing a critical role. Chromatin immunoprecipitation/ reimmunoprecipitation studies showed association of NEIL2, RNA polymerase II, and hnRNP-U on transcribed but not on transcriptionally silent genes. Furthermore, NEIL2-depleted cells accumulated more DNA damage in active than in silent genes. These results strongly support the preferential role of NEIL2 in repairing oxidized bases in the transcribed genes of mammalian cells.

UR - http://www.scopus.com/inward/record.url?scp=79953176665&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79953176665&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.198796

DO - 10.1074/jbc.M110.198796

M3 - Article

VL - 286

SP - 6006

EP - 6016

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 8

ER -