Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/helicase and a species-specific interaction between RPA and Dna2.
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 (approximately 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 delta-mediated displacement DNA synthesis are mostly short in C.elegans eukaryotes, and hence less dependent on Dna2 for viability.
Kim
Do-Hyung
DH
National Creative Research Initiative Center for Cell Cycle Control, Department of Biological Sciences, Korea Advanced Institute of Science and Technology Daejeon 305-701, Korea.
Lee
Kyoung-Hwa
KH
Kim
Jeong-Hoon
JH
Ryu
Gi-Hyuck
GH
Bae
Sung-Ho
SH
Lee
Byung-Chul
BC
Moon
Kyeong-Yeop
KY
Byun
Si-Myung
SM
Koo
Hyeon-Sook
HS
Seo
Yeon-Soo
YS
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D012313:RNA / Q000737:chemistry; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013045:Species Specificity
D029742:Caenorhabditis elegans Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D051716:Replication Protein A; D029701:Saccharomyces cerevisiae Proteins; D012313:RNA; D004706:Endodeoxyribonucleases; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; C410222:DNA2 protein, S cerevisiae; C497831:dna-2 protein, C elegans
J Biol Chem. 2002 Jul 19;277(29):26632-41
12004053
Mol Cell Biol. 2002 Jun;22(12):4202-17
12024033
J Biol Chem. 2003 Jan 17;278(3):1626-33
12424237
J Biol Chem. 2003 Jan 17;278(3):1618-25
12424238
Mol Cells. 2003 Feb 28;15(1):81-6
12661765
J Biol Chem. 2003 May 2;278(18):15983-90
12473672
Nucleic Acids Res. 2003 Jun 15;31(12):3006-15
12799426
Proc Natl Acad Sci U S A. 2003 Jul 8;100(14):8193-8
12826610
Mol Cell Biol. 2003 Aug;23(15):5346-53
12861020
Trends Biochem Sci. 2003 Jul;28(7):384-90
12878006
Mol Gen Genet. 1976 Jan 16;143(2):119-29
765749
J Theor Biol. 1978 Apr 6;71(3):339-46
642534
J Biol Chem. 1988 Dec 25;263(36):19723-33
2848839
J Virol. 1990 Dec;64(12):5912-8
2173773
Mol Cell Biol. 1991 May;11(5):2350-61
1673224
Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):579-83
8290566
J Biol Chem. 1994 Apr 15;269(15):11121-32
8157639
Nature. 1994 May 19;369(6477):207-12
7910375
Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9803-7
7524089
Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7642-6
7644470
J Biol Chem. 1995 Nov 10;270(45):26766-9
7592912
Genomics. 1996 Nov 1;37(3):408-10
8938459
J Biol Chem. 1997 Feb 21;272(8):4647-50
9081985
Proc Natl Acad Sci U S A. 1997 Jul 8;94(14):7487-92
9207118
Mol Biol Cell. 1997 Dec;8(12):2519-37
9398673
Annu Rev Biochem. 1998;67:721-51
9759502
J Biol Chem. 1998 Oct 9;273(41):26880-90
9756935
Genetics. 1999 Apr;151(4):1459-70
10101169
J Biol Chem. 2000 Jan 21;275(3):1615-24
10636853
J Biol Chem. 2000 Jun 2;275(22):16518-29
10748138
Genetics. 2000 Jul;155(3):1055-67
10880469
Nucleic Acids Res. 2000 Aug 1;28(15):2873-81
10908349
J Biol Chem. 2000 Dec 1;275(48):38022-31
10984490
Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5122-7
11309502
Nucleic Acids Res. 2001 Jul 15;29(14):3069-79
11452032
Nature. 2001 Jul 26;412(6845):456-61
11473323
Curr Biol. 2001 Oct 16;11(20):R842-4
11676941
Mol Cells. 2001 Oct 31;12(2):149-57
11710514
Proc Natl Acad Sci U S A. 2001 Dec 4;98(25):14298-303
11724925
Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9924-9
12119409