Paths from DNA damage and signaling to genome rearrangements via homologous recombination

Nickoloff, Jac A.

doi:10.1016/j.mrfmmm.2017.07.008

Cited by 20 publications

(13 citation statements)

References 203 publications

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“…During DDR, phosphorylation of the amino acid residue at position 1981 of ATM is induced (Cremona & Behrens, ; Guleria & Chandna, ). After DSB, ATM is recruited to the site of DNA damage and activated via phosphorylation, converting H2AX into γH2AX (Nickoloff, ). ATM itself can recruit additional HR pathway DNA damage repair proteins, such as MRE, NBS1, and RAD51, to form the MRN complex, which promotes the HR pathway repair process (Lavin et al ., ; Zhou et al ., ).…”

Section: Discussionmentioning

confidence: 99%

Preliminary study of the homologous recombination repair pathway in mouse spermatogonial stem cells

et al. 2018

Andrology

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The present study was designed to detect DNA repair response through the homologous recombination pathway in mouse spermatogonial stem cells. Mouse spermatogonial stem cells (mSSCs) were obtained from the adult DBA/2 mouse testes by MACS sorting. mSSCs and mice animals were divided into four groups (30 min, 2, 24 h, control) and treated with ionizing irradiation while the control group received pseudo-irradiation. Proteins involved in the homologous recombination pathway (γH2AX, ATM, RAD51, CtIP, and RPA2) were assessed in mSSCs both in vitro and in vivo. Moreover, the non-homologous end-joining or homologous recombination (NHEJ/HR) reporter plasmids were transfected into mSSCs to assess NHEJ/HR pathway activity after DNA double-strand break (DSB). γH2AX, a classical DNA DSB marker, was absent in mSSCs both in vivo and in vitro after DSB repair, but was highly expressed in other tissue stem cells. In addition, ATM and phosphorylated ATM (p-ATM) were involved in DNA damage response (DDR) in mSSCs. p-ATM foci were overexpressed immediately after irradiation (30 min and 2 h), but gradually decreased over the repair time. The HR pathway-related proteins, CtIP and RPA2 were negatively regulated after treatment in Western blot (WB). NHEJ/HR reporter plasmid transfection indicated that the HR pathway played a minor role in mSSCs during DDR, consistent with the WB findings. This study demonstrates that mSSCs may have a unique response to DNA damage since crucial proteins involved in HR pathway were negatively regulated after DSB. In addition, the expression level of p-ATM, but not γH2AX, was increased after DSB, suggesting that DNA damage repair in mSSCs might be a γH2AX-independent response. Furthermore, the HR pathway may play a minor role during DDR in mSSCs.

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Section: Discussionmentioning

confidence: 99%

Preliminary study of the homologous recombination repair pathway in mouse spermatogonial stem cells

et al. 2018

Andrology

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“…Replication stress induces DSB formation and ssDNA gaps which lead to chromosomal fragility and gap formation at metaphase [54]. Increased HR activity is associated with increased sister-chromatid exchanges and recombination events (reviewed in [69]).…”

Section: Plos Onementioning

confidence: 99%

Replication stress and FOXM1 drive radiation induced genomic instability and cell transformation

Doetsch

et al. 2020

PLoS ONE

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In contrast to the vast majority of research that has focused on the immediate effects of ionizing radiation, this work concentrates on the molecular mechanism driving delayed effects that emerge in the progeny of the exposed cells. We employed functional protein arrays to identify molecular changes induced in a human bronchial epithelial cell line (HBEC3-KT) and osteosarcoma cell line (U2OS) and evaluated their impact on outcomes associated with radiation induced genomic instability (RIGI) at day 5 and 7 post-exposure to a 2Gy X-ray dose, which revealed replication stress in the context of increased FOXM1b expression. Irradiated cells had reduced DNA replication rate detected by the DNA fiber assay and increased DNA resection detected by RPA foci and phosphorylation. Irradiated cells increased utilization of homologous recombination-dependent repair detected by a gene conversion assay and DNA damage at mitosis reflected by RPA positive chromosomal bridges, micronuclei formation and 53BP1 positive bodies in G1, all known outcomes of replication stress. Interference with the function of FOXM1, a transcription factor widely expressed in cancer, employing an aptamer, decreased radiation-induced micronuclei formation and cell transformation while plasmid-driven overexpression of FOXM1b was sufficient to induce replication stress, micronuclei formation and cell transformation.

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“…Under these circumstances, E1–E2 replication would not be able to resolve the chicken foot structure. To continue with replication, one option is to carry out break-induced replication (BIR) [ 78 , 79 , 80 ]. Using this process, a DSB is introduced into the stalled fork, allowing the restart of replication from the DSB.…”

Section: Using the Hpv16 Replication Assay To Study Lesions In Hosmentioning

confidence: 99%

Using a Human Papillomavirus Model to Study DNA Replication and Repair of Wild Type and Damaged DNA Templates in Mammalian Cells

Das

Bristol

Pichierri

et al. 2020

IJMS

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Human papillomaviruses have 8kbp DNA episomal genomes that replicate autonomously from host DNA. During initial infection, the virus increases its copy number to 20–50 copies per cell, causing torsional stress on the replicating DNA. This activates the DNA damage response (DDR) and HPV replicates its genome, at least in part, using homologous recombination. An active DDR is on throughout the HPV life cycle. Two viral proteins are required for replication of the viral genome; E2 binds to 12bp palindromic sequences around the A/T rich origin of replication and recruits the viral helicase E1 via a protein–protein interaction. E1 forms a di-hexameric complex that replicates the viral genome in association with host factors. Transient replication assays following transfection with E1–E2 expression plasmids, along with an origin containing plasmid, allow monitoring of E1-E2 replication activity. Incorporating a bacterial lacZ gene into the origin plasmid allows for the determination of replication fidelity. Here we describe how we exploited this system to investigate replication and repair in mammalian cells, including using damaged DNA templates. We propose that this system has the potential to enhance the understanding of cellular components involved in DNA replication and repair.

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Paths from DNA damage and signaling to genome rearrangements via homologous recombination

Cited by 20 publications

References 203 publications

Preliminary study of the homologous recombination repair pathway in mouse spermatogonial stem cells

Preliminary study of the homologous recombination repair pathway in mouse spermatogonial stem cells

Replication stress and FOXM1 drive radiation induced genomic instability and cell transformation

Using a Human Papillomavirus Model to Study DNA Replication and Repair of Wild Type and Damaged DNA Templates in Mammalian Cells

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