The Trans-autostimulatory Activity of Rad27 Suppresses dna2 Defects in Okazaki Fragment Processing

Munashingha, Palinda Ruvan; Lee, Chul-Hwan; Kang, Young‐Hoon; Shin, Yong-Keol; Nguyen, Tuan Anh; Seo, Yeon‐Soo

doi:10.1074/jbc.m111.326470

Cited by 8 publications

(17 citation statements)

References 50 publications

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“…B, lanes 8–10). The failure of GST‐H4N to stimulate Rad27Δ16C suggests that the N‐terminal tail of histone H4 stimulates Rad27 in a similar way to the polybasic C‐terminal tail of Rad27; the unstructured and polybasic C‐terminus of Rad27 is essential for trans ‐autostimulation of Rad27 activity . The control GST protein (G) did not show any stimulatory activity (Fig.…”

Section: Resultsmentioning

confidence: 94%

“…However, there are many other possible explanations because of the multiple known roles played by the C‐terminal tail of Rad27. For example, the C‐terminal 16‐aa tail of Rad27 is sufficient and necessary for trans ‐autostimulation , and many DNA repair factors interact with hFen1 via the C terminus . Thus, the defects of rad27Δ16C mutant we mentioned above could be in part attributed to the failure of Rad27Δ16C to interact with other factors.…”

Section: Discussionmentioning

confidence: 94%

“…Thus, the defects of rad27Δ16C mutant we mentioned above could be in part attributed to the failure of Rad27Δ16C to interact with other factors. Equally noteworthy is the importance of the C terminus of Fen1 in binding to substrate DNA in vitro and its down‐regulation by p300‐catalyzed acetylation of lysine residues in human cells . As it is not likely that recruitment of Fen1 to chromatin relies on exclusively the intrinsic DNA‐binding activity of Fen1 C terminus or its ability to interact with histones, the phenotypes of rad27Δ16C mutant observed could be due to both processes.…”

Section: Discussionmentioning

confidence: 99%

“…For example, overexpression of Rad27 suppressed temperature‐sensitive growth defects of dna2 mutant alleles, and Dna2 and Fen1 were coimmunoprecipitated . Recently, it was reported that physical interactions between Dna2 and Rad27 led to marked mutual stimulation of nuclease activities . Thus, it appears that this functional interaction between the two enzymes could contribute to rapid removal of the aberrant DNA intermediates such as 5′ or double flaps more effectively than otherwise.…”

Section: Introductionmentioning

confidence: 99%

“…Thus, it appears that this functional interaction between the two enzymes could contribute to rapid removal of the aberrant DNA intermediates such as 5′ or double flaps more effectively than otherwise. More interestingly, it was shown that Rad27 was trans ‐autostimulatory, accounting for its robust nuclease activity; the short C‐terminal tail of Rad27 was sufficient and necessary for the trans ‐autostimulation in a manner dependent on the positively charged residues in the tail , which is intrinsically unstructured . Both polybasicity and unstructured nature of the C‐terminal tail of Rad27 are reminiscent of the characteristic features of the N‐terminal tails of histones .…”

Section: Introductionmentioning

confidence: 99%

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Physical and functional interactions between nucleosomes and Rad27, a critical component of DNA processing during DNA metabolism

et al. 2016

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Highly conserved eukaryotic histones are polybasic proteins that package DNA into nucleosomes, a building block of chromatin, allowing extremely long DNA molecules to form compact and discrete chromosomes. The histone N-terminal tails that extend from the nucleosome core act as docking sites for many proteins through diverse post-translational modifications, regulating various DNA transactions. In this report, we present evidence that the nucleosomes can positively regulate the enzymatic activity of Rad27 (yeast Fen1), a major processing enzyme important for Okazaki fragment in eukaryotes. We found that individual histones, histone octamers, and nucleosomes are able to stimulate Rad27 in a manner dependent on the N-terminal tails of histones. Kinetic analyses suggest that an increase in catalytic efficiency of Rad27 was mainly due to increased affinity between DNA substrates and Rad27. It appears that the physical interaction in vivo between histones and Rad27 results in the enrichment of Rad27 in the vicinity of chromatin, increasing the availability of Rad27 for various DNA metabolisms. These results indicate that nucleosomes are not a mere structural component of chromatin, but an active regulator of DNA metabolisms that serves to ensure the efficient and faithful processing of structural intermediates arising during DNA transactions.

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Section: Resultsmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Physical and functional interactions between nucleosomes and Rad27, a critical component of DNA processing during DNA metabolism

et al. 2016

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Rad52/Rad59-dependent Recombination as a Means to Rectify Faulty Okazaki Fragment Processing

Lee

Demin

et al. 2014

Journal of Biological Chemistry

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Background: How faulty Okazaki fragments are repaired remains unclear. Results: The DNA annealing activity of Rad52 and sister chromatid cohesion are important in the repair of faulty Okazaki fragments. Conclusion: Rad52/Rad59-mediated sister chromatid recombination is a major means of repairing faulty Okazaki fragments. Significance: The faithful repair of faulty Okazaki fragments is critical for genome stability.

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Flap endonuclease Rad27 cleaves the RNA of R-loop structures to suppress telomere recombination

Liu

Chan

et al. 2023

Nucleic Acids Research

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The long non-coding telomeric RNA transcript TERRA, in the form of an RNA–DNA duplex, regulates telomere recombination. In a screen for nucleases that affects telomere recombination, mutations in DNA2, EXO1, MRE11 and SAE2 cause severe delay in type II survivor formation, indicating that type II telomere recombination is mediated through a mechanism similar to repairing double-strand breaks. On the other hand, mutation in RAD27 results in early formation of type II recombination, suggesting that RAD27 acts as a negative regulator in telomere recombination. RAD27 encodes a flap endonuclease that plays a role in DNA metabolism, including replication, repair and recombination. We demonstrate that Rad27 suppresses the accumulation of the TERRA-associated R-loop and selectively cleaves TERRA of R-loop and double-flapped structures in vitro. Moreover, we show that Rad27 negatively regulates single-stranded C-rich telomeric DNA circles (C-circles) in telomerase-deficient cells, revealing a close correlation between R-loop and C-circles during telomere recombination. These results demonstrate that Rad27 participates in telomere recombination by cleaving TERRA in the context of an R-loop or flapped RNA–DNA duplex, providing mechanistic insight into how Rad27 maintains chromosome stability by restricting the accumulation of the R-loop structure within the genome.

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The Trans-autostimulatory Activity of Rad27 Suppresses dna2 Defects in Okazaki Fragment Processing

Cited by 8 publications

References 50 publications

Physical and functional interactions between nucleosomes and Rad27, a critical component of DNA processing during DNA metabolism

Physical and functional interactions between nucleosomes and Rad27, a critical component of DNA processing during DNA metabolism

Rad52/Rad59-dependent Recombination as a Means to Rectify Faulty Okazaki Fragment Processing

Flap endonuclease Rad27 cleaves the RNA of R-loop structures to suppress telomere recombination

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