Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (416K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (16)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Bae, S.-H.
Right arrow Articles by Seo, Y.-S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bae, S.-H.
Right arrow Articles by Seo, Y.-S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 14 3069-3079
© 2001 Oxford University Press

Tripartite structure of Saccharomyces cerevisiae Dna2 helicase/endonuclease

Sung-Ho Bae, Jung-Ae Kim, Eunju Choi, Kyoung-Hwa Lee, Ho-Young Kang, Hee-Dai Kim, Jae-Hoon Kim, Kwang-Hee Bae, Yunje Cho1, Chankyu Park2 and Yeon-Soo Seo*

National Creative Research Initiative Center for Cell Cycle Control, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, 300 Chunchun-Dong, Changan-Ku, Suwon, Kyunggi-Do 440-746, Korea, 1Department of Life Science, and Division of Molecular and Life Science, Pohang University of Science and Technology, Pohang, Kyungbuk 790-784, Korea and 2Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yusung-Ku, Taejon 305-701, Korea

In order to gain insights into the structural basis of the multifunctional Dna2 enzyme involved in Okazaki fragment processing, we performed biochemical, biophysical and genetic studies to dissect the domain structure of Dna2. Proteolytic digestion of Dna2 using subtilisin produced a 127 kDa polypeptide that lacked the 45 kDa N-terminal region of Dna2. Further digestion generated two subtilisin-resistant core fragments of approximately equal size, 58 and 60 kDa. Surprisingly, digestion resulted in a significant (3- to 8-fold) increase in both ATPase and endonuclease activities compared to the intact enzyme. However, cells with a mutant DNA2 allele lacking the corresponding N-terminal region were severely impaired in growth, being unable to grow at 37°C, indicating that the N-terminal region contains a domain critical for a cellular function(s) of Dna2. Analyses of the hydrodynamic properties of and in vivo complex formation by wild-type and/or mutant Dna2 lacking the N-terminal 45 kDa domain revealed that Dna2 is active as the monomer and thus the defect in the mutant Dna2 protein is not due to its inability to multimerize. In addition, we found that the N-terminal 45 kDa domain interacts physically with a central region located between the two catalytic domains. Our results suggest that the N-terminal 45 kDa domain of Dna2 plays a critical role in regulation of the enzymatic activities of Dna2 by serving as a site for intra- and intermolecular interactions essential for optimal function of Dna2 in Okazaki fragment processing. The possible mode of regulation of Dna2 is discussed based upon our recent finding that replication protein A interacts functionally and physically with Dna2 during Okazaki fragment processing.

* To whom correspondence should be addressed. Tel: +82 31 299 6440; Fax: +82 31 299 6435; Email: ysseo{at}med.skku.ac.kr


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Gen. Virol.Home page
V. V. Makarov, E. N. Rybakova, A. V. Efimov, E. N. Dobrov, M. V. Serebryakova, A. G. Solovyev, I. V. Yaminsky, M. E. Taliansky, S. Yu. Morozov, and N. O. Kalinina
Domain organization of the N-terminal portion of hordeivirus movement protein TGBp1
J. Gen. Virol., December 1, 2009; 90(12): 3022 - 3032.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y.-H. Kang, M.-J. Kang, J.-H. Kim, C.-H. Lee, I.-t. Cho, J. Hurwitz, and Y.-S. Seo
The MPH1 Gene of Saccharomyces cerevisiae Functions in Okazaki Fragment Processing
J. Biol. Chem., April 17, 2009; 284(16): 10376 - 10386.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. A. Stewart, J. L. Campbell, and R. A. Bambara
Flap Endonuclease Disengages Dna2 Helicase/Nuclease from Okazaki Fragment Flaps
J. Biol. Chem., December 15, 2006; 281(50): 38565 - 38572.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J.-H. Kim, H.-D. Kim, G.-H. Ryu, D.-H. Kim, J. Hurwitz, and Y.-S. Seo
Isolation of human Dna2 endonuclease and characterization of its enzymatic properties.
Nucleic Acids Res., January 1, 2006; 34(6): 1854 - 1864.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
J.-H. Kim, Y.-H. Kang, H.-J. Kang, D.-H. Kim, G.-H. Ryu, M.-J. Kang, and Y.-S. Seo
In vivo and in vitro studies of Mgs1 suggest a link between genome instability and Okazaki fragment processing
Nucleic Acids Res., October 26, 2005; 33(19): 6137 - 6150.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
D.-H. Kim, K.-H. Lee, J.-H. Kim, G.-H. Ryu, S.-H. Bae, B.-C. Lee, K.-Y. Moon, S.-M. Byun, H.-S. Koo, and Y.-S. Seo
Enzymatic properties of the Caenorhabditis elegans Dna2 endonuclease/helicase and a species-specific interaction between RPA and Dna2
Nucleic Acids Res., March 3, 2005; 33(4): 1372 - 1383.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H.-I Kao, J. L. Campbell, and R. A. Bambara
Dna2p Helicase/Nuclease Is a Tracking Protein, Like FEN1, for Flap Cleavage during Okazaki Fragment Maturation
J. Biol. Chem., December 3, 2004; 279(49): 50840 - 50849.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
H. Tanaka, G.-H. Ryu, Y.-S. Seo, and S. A. MacNeill
Genetics of lagging strand DNA synthesis and maturation in fission yeast: suppression analysis links the Dna2-Cdc24 complex to DNA polymerase {delta}
Nucleic Acids Res., December 2, 2004; 32(21): 6367 - 6377.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
K. Tomita, T. Kibe, H.-Y. Kang, Y.-S. Seo, M. Uritani, T. Ushimaru, and M. Ueno
Fission Yeast Dna2 Is Required for Generation of the Telomeric Single-Strand Overhang
Mol. Cell. Biol., November 1, 2004; 24(21): 9557 - 9567.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
J. B. Bessler and V. A. Zakian
The Amino Terminus of the Saccharomyces cerevisiae DNA Helicase Rrm3p Modulates Protein Function Altering Replication and Checkpoint Activity
Genetics, November 1, 2004; 168(3): 1205 - 1218.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
G.-H. Ryu, H. Tanaka, D.-H. Kim, J.-H. Kim, S.-H. Bae, Y.-N. Kwon, J. S. Rhee, S. A. MacNeill, and Y.-S. Seo
Genetic and biochemical analyses of Pfh1 DNA helicase function in fission yeast
Nucleic Acids Res., August 9, 2004; 32(14): 4205 - 4216.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
K.-H. Bae, H.-S. Kim, S.-H. Bae, H.-Y. Kang, S. Brill, and Y.-S. Seo
Bimodal interaction between replication-protein A and Dna2 is critical for Dna2 function both in vivo and in vitro
Nucleic Acids Res., June 15, 2003; 31(12): 3006 - 3015.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Weitao, M. Budd, L. L. M. Hoopes, and J. L. Campbell
Dna2 Helicase/Nuclease Causes Replicative Fork Stalling and Double-strand Breaks in the Ribosomal DNA of Saccharomyces cerevisiae
J. Biol. Chem., June 13, 2003; 278(25): 22513 - 22522.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Higashibata, H. Kikuchi, Y. Kawarabayasi, and I. Matsui
Helicase and Nuclease Activities of Hyperthermophile Pyrococcus horikoshii Dna2 Inhibited by Substrates with RNA Segments at 5'-End
J. Biol. Chem., April 25, 2003; 278(18): 15983 - 15990.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S.-H. Bae, D. W. Kim, J. Kim, J.-H. Kim, D.-H. Kim, H.-D. Kim, H.-Y. Kang, and Y.-S. Seo
Coupling of DNA Helicase and Endonuclease Activities of Yeast Dna2 Facilitates Okazaki Fragment Processing
J. Biol. Chem., July 12, 2002; 277(29): 26632 - 26641.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.