Interaction of the human DNA glycosylase NEIL1 with proliferating cell nuclear antigen

The potential for replication-associated repair of oxidized bases in mammalian genomes

Hong Dou, Corey A. Theriot, Aditi Das, Muralidhar L. Hegde, Yoshihiro Matsumoto, Istvan Boldogh, Tapas K. Hazra, Kishor K Bhakat, Sankar Mitra

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

NEIL1 and NEIL2 compose a family of DNA glycosylases that is distinct from that of the other two DNA glycosylases, OGG1 and NTH1, all of which are involved in repair of oxidized bases in mammalian genomes. That the NEIL proteins, unlike OGG1 and NTH1, are able to excise base lesions from single-stranded DNA regions suggests their preferential involvement in repair during replication and/or transcription. Previous studies showing S phase-specific activation of NEIL1, but not NEIL2, suggested NEIL1 involvement in the repair of replicating DNA. Here, we show that human NEIL1 stably interacts both in vivo and in vitro with proliferating cell nuclear antigen (PCNA), the sliding clamp for DNA replication. PCNA stimulates NEIL1 activity in excising the oxidized base 5-hydroxyuracil from single-stranded DNA sequences including fork structures. PCNA enhances NEIL1 loading on the substrate. In contrast, although present in the NEIL2 immunocomplex, PCNA does not stimulate NEIL2. NEIL1 interacts with PCNA via a domain that is located in a region near the C terminus, dispensable for base excision activity. The interacting sequence in NEIL1, which lacks the canonical PCNA-binding motif, includes a sequence conserved in DNA polymerase δ and implicated in its PCNA binding. Mammalian two-hybrid analysis confirmed PCNA interaction with NEIL1. The G127A mutation in PCNA reduces its stimulatory activity, suggesting that the interdomain connector loop, a common binding interface of PCNA, is involved in NEIL1 binding. These results strongly support in vivo function of NEIL1 in preferential repair of oxidized bases inDNAprior to replication.

Original languageEnglish (US)
Pages (from-to)3130-3140
Number of pages11
JournalJournal of Biological Chemistry
Volume283
Issue number6
DOIs
StatePublished - Feb 8 2008

Fingerprint

DNA Glycosylases
Proliferating Cell Nuclear Antigen
Repair
Genes
Genome
Single-Stranded DNA
Conserved Sequence
DNA sequences
DNA
Clamping devices
DNA-Directed DNA Polymerase
Transcription
DNA Replication
S Phase
DNA Repair
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Interaction of the human DNA glycosylase NEIL1 with proliferating cell nuclear antigen : The potential for replication-associated repair of oxidized bases in mammalian genomes. / Dou, Hong; Theriot, Corey A.; Das, Aditi; Hegde, Muralidhar L.; Matsumoto, Yoshihiro; Boldogh, Istvan; Hazra, Tapas K.; Bhakat, Kishor K; Mitra, Sankar.

In: Journal of Biological Chemistry, Vol. 283, No. 6, 08.02.2008, p. 3130-3140.

Research output: Contribution to journalArticle

Dou, Hong ; Theriot, Corey A. ; Das, Aditi ; Hegde, Muralidhar L. ; Matsumoto, Yoshihiro ; Boldogh, Istvan ; Hazra, Tapas K. ; Bhakat, Kishor K ; Mitra, Sankar. / Interaction of the human DNA glycosylase NEIL1 with proliferating cell nuclear antigen : The potential for replication-associated repair of oxidized bases in mammalian genomes. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 6. pp. 3130-3140.
@article{8d02546ee80f42b7b5e83153fd3d2901,
title = "Interaction of the human DNA glycosylase NEIL1 with proliferating cell nuclear antigen: The potential for replication-associated repair of oxidized bases in mammalian genomes",
abstract = "NEIL1 and NEIL2 compose a family of DNA glycosylases that is distinct from that of the other two DNA glycosylases, OGG1 and NTH1, all of which are involved in repair of oxidized bases in mammalian genomes. That the NEIL proteins, unlike OGG1 and NTH1, are able to excise base lesions from single-stranded DNA regions suggests their preferential involvement in repair during replication and/or transcription. Previous studies showing S phase-specific activation of NEIL1, but not NEIL2, suggested NEIL1 involvement in the repair of replicating DNA. Here, we show that human NEIL1 stably interacts both in vivo and in vitro with proliferating cell nuclear antigen (PCNA), the sliding clamp for DNA replication. PCNA stimulates NEIL1 activity in excising the oxidized base 5-hydroxyuracil from single-stranded DNA sequences including fork structures. PCNA enhances NEIL1 loading on the substrate. In contrast, although present in the NEIL2 immunocomplex, PCNA does not stimulate NEIL2. NEIL1 interacts with PCNA via a domain that is located in a region near the C terminus, dispensable for base excision activity. The interacting sequence in NEIL1, which lacks the canonical PCNA-binding motif, includes a sequence conserved in DNA polymerase δ and implicated in its PCNA binding. Mammalian two-hybrid analysis confirmed PCNA interaction with NEIL1. The G127A mutation in PCNA reduces its stimulatory activity, suggesting that the interdomain connector loop, a common binding interface of PCNA, is involved in NEIL1 binding. These results strongly support in vivo function of NEIL1 in preferential repair of oxidized bases inDNAprior to replication.",
author = "Hong Dou and Theriot, {Corey A.} and Aditi Das and Hegde, {Muralidhar L.} and Yoshihiro Matsumoto and Istvan Boldogh and Hazra, {Tapas K.} and Bhakat, {Kishor K} and Sankar Mitra",
year = "2008",
month = "2",
day = "8",
doi = "10.1074/jbc.M709186200",
language = "English (US)",
volume = "283",
pages = "3130--3140",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "6",

}

TY - JOUR

T1 - Interaction of the human DNA glycosylase NEIL1 with proliferating cell nuclear antigen

T2 - The potential for replication-associated repair of oxidized bases in mammalian genomes

AU - Dou, Hong

AU - Theriot, Corey A.

AU - Das, Aditi

AU - Hegde, Muralidhar L.

AU - Matsumoto, Yoshihiro

AU - Boldogh, Istvan

AU - Hazra, Tapas K.

AU - Bhakat, Kishor K

AU - Mitra, Sankar

PY - 2008/2/8

Y1 - 2008/2/8

N2 - NEIL1 and NEIL2 compose a family of DNA glycosylases that is distinct from that of the other two DNA glycosylases, OGG1 and NTH1, all of which are involved in repair of oxidized bases in mammalian genomes. That the NEIL proteins, unlike OGG1 and NTH1, are able to excise base lesions from single-stranded DNA regions suggests their preferential involvement in repair during replication and/or transcription. Previous studies showing S phase-specific activation of NEIL1, but not NEIL2, suggested NEIL1 involvement in the repair of replicating DNA. Here, we show that human NEIL1 stably interacts both in vivo and in vitro with proliferating cell nuclear antigen (PCNA), the sliding clamp for DNA replication. PCNA stimulates NEIL1 activity in excising the oxidized base 5-hydroxyuracil from single-stranded DNA sequences including fork structures. PCNA enhances NEIL1 loading on the substrate. In contrast, although present in the NEIL2 immunocomplex, PCNA does not stimulate NEIL2. NEIL1 interacts with PCNA via a domain that is located in a region near the C terminus, dispensable for base excision activity. The interacting sequence in NEIL1, which lacks the canonical PCNA-binding motif, includes a sequence conserved in DNA polymerase δ and implicated in its PCNA binding. Mammalian two-hybrid analysis confirmed PCNA interaction with NEIL1. The G127A mutation in PCNA reduces its stimulatory activity, suggesting that the interdomain connector loop, a common binding interface of PCNA, is involved in NEIL1 binding. These results strongly support in vivo function of NEIL1 in preferential repair of oxidized bases inDNAprior to replication.

AB - NEIL1 and NEIL2 compose a family of DNA glycosylases that is distinct from that of the other two DNA glycosylases, OGG1 and NTH1, all of which are involved in repair of oxidized bases in mammalian genomes. That the NEIL proteins, unlike OGG1 and NTH1, are able to excise base lesions from single-stranded DNA regions suggests their preferential involvement in repair during replication and/or transcription. Previous studies showing S phase-specific activation of NEIL1, but not NEIL2, suggested NEIL1 involvement in the repair of replicating DNA. Here, we show that human NEIL1 stably interacts both in vivo and in vitro with proliferating cell nuclear antigen (PCNA), the sliding clamp for DNA replication. PCNA stimulates NEIL1 activity in excising the oxidized base 5-hydroxyuracil from single-stranded DNA sequences including fork structures. PCNA enhances NEIL1 loading on the substrate. In contrast, although present in the NEIL2 immunocomplex, PCNA does not stimulate NEIL2. NEIL1 interacts with PCNA via a domain that is located in a region near the C terminus, dispensable for base excision activity. The interacting sequence in NEIL1, which lacks the canonical PCNA-binding motif, includes a sequence conserved in DNA polymerase δ and implicated in its PCNA binding. Mammalian two-hybrid analysis confirmed PCNA interaction with NEIL1. The G127A mutation in PCNA reduces its stimulatory activity, suggesting that the interdomain connector loop, a common binding interface of PCNA, is involved in NEIL1 binding. These results strongly support in vivo function of NEIL1 in preferential repair of oxidized bases inDNAprior to replication.

UR - http://www.scopus.com/inward/record.url?scp=41249094475&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41249094475&partnerID=8YFLogxK

U2 - 10.1074/jbc.M709186200

DO - 10.1074/jbc.M709186200

M3 - Article

VL - 283

SP - 3130

EP - 3140

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 6

ER -