Proteomics

Dataset Information

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Monitoring of the spatial and temporal dynamics of BER/SSBR pathway proteins, including MYH, UNG2, MPG, NTH1 and NEIL1-3, during DNA replication, Batch 2


ABSTRACT: Base lesions in DNA can stall the replication machinery or induce mutations if bypassed. Consequently, lesions must be repaired before replication or in a post-replicative process to maintain genomic stability. Base excision repair (BER) is the main pathway for repair of base lesions, but how BER is organized during DNA replication is unclear. Here we refined the iPOND (isolation of proteins on nascent DNA) technique with targeted mass-spectrometry analysis, which enabled us to detect all proteins required for BER on nascent DNA and to monitor their spatiotemporal orchestration at replication forks. We demonstrate that XRCC1 and other BER/single-strand break repair (SSBR) proteins are enriched in replisomes in unstressed cells, supporting a cellular capacity of post-replicative BER/SSBR. Importantly, the DNA glycosylases MYH, UNG2, MPG, NTH1, NEIL1 and NEIL3 were also detected on nascent DNA. Thus our findings reveal that a broad spectrum of DNA base lesions are recognized and repaired by BER in a post-replicative process.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Early Embryonic Cell

SUBMITTER: Animesh Sharma  

LAB HEAD: Marit Otterlei

PROVIDER: PXD004494 | Pride | 2017-11-23

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
20160504_iPOND_15P_01.raw Raw
20160504_iPOND_15P_02.raw Raw
20160504_iPOND_15P_03.raw Raw
20160504_iPOND_5P_01.raw Raw
20160504_iPOND_5P_02.raw Raw
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Publications

Monitoring of the spatial and temporal dynamics of BER/SSBR pathway proteins, including MYH, UNG2, MPG, NTH1 and NEIL1-3, during DNA replication.

Bj Rås Karine Ø KØ   Sousa Mirta M L MML   Sharma Animesh A   Fonseca Davi M DM   S Gaard Caroline K CK   Bj Rås Magnar M   Otterlei Marit M  

Nucleic acids research 20170801 14


Base lesions in DNA can stall the replication machinery or induce mutations if bypassed. Consequently, lesions must be repaired before replication or in a post-replicative process to maintain genomic stability. Base excision repair (BER) is the main pathway for repair of base lesions and is known to be associated with DNA replication, but how BER is organized during replication is unclear. Here we coupled the iPOND (isolation of proteins on nascent DNA) technique with targeted mass-spectrometry  ...[more]

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