Nek1 Regulates Rad54 to Orchestrate Homologous Recombination and Replication Fork Stability

SUMMARY Inaccurate repair of broken chromosomes generates structural variants that can fuel evolution and inflict pathology. We describe a novel rearrangement mechanism in which translocation between intact chromosomes is induced by a lesion on a third chromosome. This multi-invasion-induced rearrangement (MIR) stems from a homologous recombination byproduct, where a broken DNA end simultaneously invades two intact donors. No homology is required between the donors, and the intervening sequence from the invading molecule is inserted at the translocation site. MIR is stimulated by increasing homology length and spatial proximity of the donors, and depends on the overlapping activities of the structure-selective endonucleases Mus81-Mms4, Slx1-Slx4, and Yen1. Conversely, the 3′-flap nuclease Rad1-Rad10 and enzymes known to disrupt recombination intermediates (Sgs1-Top3-Rmi1, Srs2 and Mph1) inhibit MIR. Resolution of MIR intermediates propagates secondary chromosome breaks that frequently cause additional rearrangements. MIR features have implications for the formation of simple and complex rearrangements underlying human pathologies.

show abstract

“…Human RAD51 and RPA were purified as described (Sneeden et al, 2013). Human RAD54 was purified as described (Spies et al, 2016). …”

Section: Star Methodsmentioning

confidence: 99%

Multi-invasions Are Recombination Byproducts that Induce Chromosomal Rearrangements

Piazza

Wright

Heyer

2017

Cell

Self Cite

112

189

View full text Add to dashboard Cite

show abstract

“…Nucleolytic resection following replication fork stalling can be monitored using the same labeling scheme described above [13, 14, 16, 19, 42, 43]. In this case, nucleolytic resection of both strands of a stalled replication forks results in shortening of the first tract (the one where the thymidine analog was incorporated before drug treatment) (Figure 2C).…”

Section: Single Molecule Dna Fibermentioning

confidence: 99%

Combining electron microscopy with single molecule DNA fiber approaches to study DNA replication dynamics

Vindigni

Lopes

2017

Biophysical Chemistry

View full text Add to dashboard Cite

Summary Replication stress is a crucial driver of genomic instability. Understanding the mechanisms of replication stress response is instrumental to improve diagnosis and treatment of human disease. Electron microscopy (EM) is currently the only technique that allows to directly visualize a high number of replication intermediates and to monitor their remodeling upon stress. At the same time, DNA fiber analysis is useful to gain mechanistic insight on how genotoxic agents perturb replication fork dynamics genome-wide at single-molecule resolution. Combining these techniques has proven invaluable to achieve a comprehensive view of the mechanisms that ensure error-free processing of damaged replication forks. Here, we review how EM and single-molecule DNA fiber approaches can be used together to shed light into the mechanisms of replication stress response and discuss important cautions to be taken into account when comparing results obtained by EM and DNA fiber.

show abstract

“…As verified in recent work, the RAD51AP1-UAF1 complex promotes RAD51-mediated homologous DNA pairing [35]; NEK1 can orchestrate homologous recombination and replication fork stability by regulating RAD54 [36]; and MAPK7 plays a key role in controlling cell proliferation, migration, and invasion in osteosarcoma [37]. We also performed further experiments to validate the phenotypic changes in NSCs caused by SIRT1 deletion, with neurosphere formation and Ki67 immunostaining results demonstrating the inactivating role of SIRT1 in proliferation and γH2AX staining data revealing the need for SIRT1 in DNA damage repair.…”

Section: Cellular Physiology and Biochemistrymentioning

confidence: 56%

SIRT1 Involved in the Regulation of Alternative Splicing Affects the DNA Damage Response in Neural Stem Cells

Wang

Ren

et al. 2018

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/Aims: Alternative splicing and DNA damage exhibit cross-regulation, with not only DNA damage inducing changes in alternative splicing, but alternative splicing itself possibly modulating the DNA damage response (DDR). Sirt1, a prominent anti-aging player, plays pivotal roles in the DDR. However, few studies have examined alternative splicing with DNA damage in neural stem cells (NSCs) and, in essence, nothing is known about whether SIRT1 regulates alternative splicing. Hence, we investigated the potential involvement of Sirt1-mediated alternative splicing in the NSC DDR. Methods: Genome-wide alternative splicing profiling was performed upon DNA damage induction and SIRT1 deletion. Results: DNA damage caused genome-wide changes in alternative splicing in adult NSCs and Sirt1 deficiency dramatically altered DDR-related alternative splicing. In particular, extensive alternative splicing changes in DDR-related processes such as cell cycle control and DNA damage repair were observed; these processes were dramatically influenced by Sirt1 deficiency. Phenotypically, Sirt1 deficiency altered the proliferation and DNA repair of adult NSCs, possibly by regulating alternative splicing. Conclusion: SIRT1 helps to regulate alternative splicing, which itself affects the DDR of NSCs. Our findings provide novel insight into the mechanisms underlying the DDR in stem cells.

show abstract

Nek1 Regulates Rad54 to Orchestrate Homologous Recombination and Replication Fork Stability

Cited by 84 publications

References 58 publications

Multi-invasions Are Recombination Byproducts that Induce Chromosomal Rearrangements

Multi-invasions Are Recombination Byproducts that Induce Chromosomal Rearrangements

Combining electron microscopy with single molecule DNA fiber approaches to study DNA replication dynamics

SIRT1 Involved in the Regulation of Alternative Splicing Affects the DNA Damage Response in Neural Stem Cells

Contact Info

Product

Resources

About