Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/ helicase and a species-specific interaction between RPA and Dna2

Do Hyung Kim, Kyoung Hwa Lee, Jeong Hoon Kim, Gi Hyuck Ryu, Sung Ho Bae, Cyung Chul Lee, Kyeong Yeop Moon, Si Myung Byun, Hyeon Sook Koo, Yeon Soo Seo

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In both budding and fission yeasts, a null mutation of the DNA2 gene is lethal. In contrast, a null mutation of Caenorhabditis elegans dna2+ causes a delayed lethality, allowing survival of some mutant C.elegans adults to F2 generation. In order to understand reasons for this difference in requirement of Dna2 between these organisms, we examined the enzymatic properties of the recombinant C.elegans Dna2 (CeDna2) and its interaction with replication-protein A (RPA) from various sources. Like budding yeast Dna2, CeDna2 possesses DNA-dependent ATPase, helicase and endonuclease activities. The specific activities of both ATPase and endonuclease activities of the CeDna2 were considerably higher than the yeast Dna2 (∼10- and 20-fold, respectively). CeDna2 endonuclease efficiently degraded a short 5′ single-stranded DNA tail (<10 nt) that was hardly cleaved by ScDna2. Both endonuclease and helicase activities of CeDna2 were stimulated by CeRPA, but not by human or yeast RPA, demonstrating a species-specific interaction between Dna2 and RPA. These and other enzymatic properties of CeDna2 described in this paper may shed light on the observation that C. elegans is less stringently dependent on Dna2 for its viability than Saccharomyces cerevisiae. We propose that flaps generated by DNA polymerase δ-mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability.

Original languageEnglish
Pages (from-to)1372-1383
Number of pages12
JournalNucleic acids research
Volume33
Issue number4
DOIs
Publication statusPublished - 2005 Oct 20

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Replication Protein A
Endonucleases
Caenorhabditis elegans
Saccharomycetales
Lethal Genes
Mutation
Fungal Proteins
Schizosaccharomyces
Single-Stranded DNA
DNA-Directed DNA Polymerase
Eukaryota
Saccharomyces cerevisiae
Tail
Yeasts
Survival
DNA
DNA-dependent ATPase-endonuclease

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Kim, Do Hyung ; Lee, Kyoung Hwa ; Kim, Jeong Hoon ; Ryu, Gi Hyuck ; Bae, Sung Ho ; Lee, Cyung Chul ; Moon, Kyeong Yeop ; Byun, Si Myung ; Koo, Hyeon Sook ; Seo, Yeon Soo. / Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/ helicase and a species-specific interaction between RPA and Dna2. In: Nucleic acids research. 2005 ; Vol. 33, No. 4. pp. 1372-1383.
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abstract = "In both budding and fission yeasts, a null mutation of the DNA2 gene is lethal. In contrast, a null mutation of Caenorhabditis elegans dna2+ causes a delayed lethality, allowing survival of some mutant C.elegans adults to F2 generation. In order to understand reasons for this difference in requirement of Dna2 between these organisms, we examined the enzymatic properties of the recombinant C.elegans Dna2 (CeDna2) and its interaction with replication-protein A (RPA) from various sources. Like budding yeast Dna2, CeDna2 possesses DNA-dependent ATPase, helicase and endonuclease activities. The specific activities of both ATPase and endonuclease activities of the CeDna2 were considerably higher than the yeast Dna2 (∼10- and 20-fold, respectively). CeDna2 endonuclease efficiently degraded a short 5′ single-stranded DNA tail (<10 nt) that was hardly cleaved by ScDna2. Both endonuclease and helicase activities of CeDna2 were stimulated by CeRPA, but not by human or yeast RPA, demonstrating a species-specific interaction between Dna2 and RPA. These and other enzymatic properties of CeDna2 described in this paper may shed light on the observation that C. elegans is less stringently dependent on Dna2 for its viability than Saccharomyces cerevisiae. We propose that flaps generated by DNA polymerase δ-mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability.",
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Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/ helicase and a species-specific interaction between RPA and Dna2. / Kim, Do Hyung; Lee, Kyoung Hwa; Kim, Jeong Hoon; Ryu, Gi Hyuck; Bae, Sung Ho; Lee, Cyung Chul; Moon, Kyeong Yeop; Byun, Si Myung; Koo, Hyeon Sook; Seo, Yeon Soo.

In: Nucleic acids research, Vol. 33, No. 4, 20.10.2005, p. 1372-1383.

Research output: Contribution to journalArticle

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AU - Kim, Do Hyung

AU - Lee, Kyoung Hwa

AU - Kim, Jeong Hoon

AU - Ryu, Gi Hyuck

AU - Bae, Sung Ho

AU - Lee, Cyung Chul

AU - Moon, Kyeong Yeop

AU - Byun, Si Myung

AU - Koo, Hyeon Sook

AU - Seo, Yeon Soo

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AB - In both budding and fission yeasts, a null mutation of the DNA2 gene is lethal. In contrast, a null mutation of Caenorhabditis elegans dna2+ causes a delayed lethality, allowing survival of some mutant C.elegans adults to F2 generation. In order to understand reasons for this difference in requirement of Dna2 between these organisms, we examined the enzymatic properties of the recombinant C.elegans Dna2 (CeDna2) and its interaction with replication-protein A (RPA) from various sources. Like budding yeast Dna2, CeDna2 possesses DNA-dependent ATPase, helicase and endonuclease activities. The specific activities of both ATPase and endonuclease activities of the CeDna2 were considerably higher than the yeast Dna2 (∼10- and 20-fold, respectively). CeDna2 endonuclease efficiently degraded a short 5′ single-stranded DNA tail (<10 nt) that was hardly cleaved by ScDna2. Both endonuclease and helicase activities of CeDna2 were stimulated by CeRPA, but not by human or yeast RPA, demonstrating a species-specific interaction between Dna2 and RPA. These and other enzymatic properties of CeDna2 described in this paper may shed light on the observation that C. elegans is less stringently dependent on Dna2 for its viability than Saccharomyces cerevisiae. We propose that flaps generated by DNA polymerase δ-mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability.

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