JBC PeproTech; Our Business is Cytokines!

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M110662200 on February 1, 2002

J. Biol. Chem., Vol. 277, Issue 17, 14379-14389, April 26, 2002
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
277/17/14379    most recent
M110662200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kao, H.-I
Right arrow Articles by Bambara, R. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kao, H.-I
Right arrow Articles by Bambara, R. A.

Cleavage Specificity of Saccharomyces cerevisiae Flap Endonuclease 1 Suggests a Double-Flap Structure as the Cellular Substrate*

Hui-I Kao, Leigh A. HenricksenDagger , Yuan Liu, and Robert A. Bambara§

From the Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642

Flap endonuclease 1 (FEN1) is a structure-specific nuclease that cleaves substrates containing unannealed 5'-flaps during Okazaki fragment processing. Cleavage removes the flap at or near the point of annealing. The preferred substrate for archaeal FEN1 or the 5'-nuclease domains of bacterial DNA polymerases is a double-flap structure containing a 3'-tail on the upstream primer adjacent to the 5'-flap. We report that FEN1 in Saccharomyces cerevisiae (Rad27p) exhibits a similar specificity. Cleavage was most efficient when the upstream primer contained a 1-nucleotide 3'-tail as compared with the fully annealed upstream primer traditionally tested. The site of cleavage was exclusively at a position one nucleotide into the annealed region, allowing human DNA ligase I to seal all resulting nicks. In contrast, a portion of the products from traditional flap substrates is not ligated. The 3'-OH of the upstream primer is not critical for double-flap recognition, because Rad27p is tolerant of modifications. However, the positioning of the 3'-nucleotide defines the site of cleavage. We have tested substrates having complementary tails that equilibrate to many structures by branch migration. FEN1 only cleaved those containing a 1-nucleotide 3'-tail. Equilibrating substrates containing 12-ribonucleotides at the end of the 5'-flap simulates the situation in vivo. Rad27p cleaves this substrate in the expected 1-nucleotide 3'-tail configuration. Overall, these results suggest that the double-flap substrate is formed and cleaved during eukaryotic DNA replication in vivo.

* This work was supported by National Institutes of Health Grant GM24441.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Present address: Center of Aging and Developmental Biology, University of Rochester, 601 Elmwood Ave., Box 645, Rochester, NY 14642.

§ To whom correspondence should be addressed: Dept. of Biochemistry and Biophysics, University of Rochester Medical Center, 601 Elmwood Ave., Box 712, Rochester, NY 14642. Tel.: 585-275-3269; Fax: 585-271-2683; E-mail: robert_bambara@urmc.rochester.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
M. Hohl, I. Dunand-Sauthier, L. Staresincic, P. Jaquier-Gubler, F. Thorel, M. Modesti, S. G. Clarkson, and O. D. Scharer
Domain swapping between FEN-1 and XPG defines regions in XPG that mediate nucleotide excision repair activity and substrate specificity
Nucleic Acids Res., May 14, 2007; 35(9): 3053 - 3063.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Singh, L. Zheng, V. Chavez, J. Qiu, and B. Shen
Concerted Action of Exonuclease and Gap-dependent Endonuclease Activities of FEN-1 Contributes to the Resolution of Triplet Repeat Sequences (CTG)n- and (GAA)n-derived Secondary Structures Formed during Maturation of Okazaki Fragments
J. Biol. Chem., February 9, 2007; 282(6): 3465 - 3477.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Masuda-Sasa, P. Polaczek, and J. L. Campbell
Single Strand Annealing and ATP-independent Strand Exchange Activities of Yeast and Human DNA2: POSSIBLE ROLE IN OKAZAKI FRAGMENT MATURATION
J. Biol. Chem., December 15, 2006; 281(50): 38555 - 38564.
[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
I. Ivanov, B. R. Chapados, J. A. McCammon, and J. A. Tainer
Proliferating cell nuclear antigen loaded onto double-stranded DNA: dynamics, minor groove interactions and functional implications
Nucleic Acids Res., November 6, 2006; 34(20): 6023 - 6033.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. L. Rossi and R. A. Bambara
Reconstituted Okazaki Fragment Processing Indicates Two Pathways of Primer Removal
J. Biol. Chem., September 8, 2006; 281(36): 26051 - 26061.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
M. E. Budd, C. C. Reis, S. Smith, K. Myung, and J. L. Campbell
Evidence Suggesting that Pif1 Helicase Functions in DNA Replication with the Dna2 Helicase/Nuclease and DNA Polymerase {delta}.
Mol. Cell. Biol., April 1, 2006; 26(7): 2490 - 2500.
[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
S. Sharma, J. A. Sommers, R. K. Gary, E. Friedrich-Heineken, U. Hubscher, and R. M. Brosh Jr
The interaction site of Flap Endonuclease-1 with WRN helicase suggests a coordination of WRN and PCNA
Nucleic Acids Res., December 2, 2005; 33(21): 6769 - 6781.
[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
 GeneticsHome page
L. Li, K. M. Murphy, U. Kanevets, and L. J. Reha-Krantz
Sensitivity to Phosphonoacetic Acid: A New Phenotype to Probe DNA Polymerase {delta} in Saccharomyces cerevisiae
Genetics, June 1, 2005; 170(2): 569 - 580.
[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
 Proc. Natl. Acad. Sci. USAHome page
W. Wang, P. Brandt, M. L. Rossi, L. Lindsey-Boltz, V. Podust, E. Fanning, A. Sancar, and R. A. Bambara
The human Rad9-Rad1-Hus1 checkpoint complex stimulates flap endonuclease 1
PNAS, November 30, 2004; 101(48): 16762 - 16767.
[Abstract] [Full Text] [PDF]

Home page
 Hum Mol GenetHome page
S. Sharma, J. A. Sommers, and R. M. Brosh Jr
In vivo function of the conserved non-catalytic domain of Werner syndrome helicase in DNA replication
Hum. Mol. Genet., October 1, 2004; 13(19): 2247 - 2261.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Qiu, R. Liu, B. R. Chapados, M. Sherman, J. A. Tainer, and B. Shen
Interaction Interface of Human Flap Endonuclease-1 with Its DNA Substrates
J. Biol. Chem., June 4, 2004; 279(23): 24394 - 24402.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
E. Friedrich-Heineken and U. Hubscher
The Fen1 extrahelical 3'-flap pocket is conserved from archaea to human and regulates DNA substrate specificity
Nucleic Acids Res., May 6, 2004; 32(8): 2520 - 2528.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
Y. Liu, H. Zhang, J. Veeraraghavan, R. A. Bambara, and C. H. Freudenreich
Saccharomyces cerevisiae Flap Endonuclease 1 Uses Flap Equilibration To Maintain Triplet Repeat Stability
Mol. Cell. Biol., May 1, 2004; 24(9): 4049 - 4064.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
E. Matsui, J. Abe, H. Yokoyama, and I. Matsui
Aromatic Residues Located Close to the Active Center Are Essential for the Catalytic Reaction of Flap Endonuclease-1 from Hyperthermophilic Archaeon Pyrococcus horikoshii
J. Biol. Chem., April 16, 2004; 279(16): 16687 - 16696.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H.-I Kao, J. Veeraraghavan, P. Polaczek, J. L. Campbell, and R. A. Bambara
On the Roles of Saccharomyces cerevisiae Dna2p and Flap Endonuclease 1 in Okazaki Fragment Processing
J. Biol. Chem., April 9, 2004; 279(15): 15014 - 15024.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
S. Sharma, M. Otterlei, J. A. Sommers, H. C. Driscoll, G. L. Dianov, H.-I Kao, R. A. Bambara, and R. M. Brosh Jr.
WRN Helicase and FEN-1 Form a Complex upon Replication Arrest and Together Process Branchmigrating DNA Structures Associated with the Replication Fork
Mol. Biol. Cell, February 1, 2004; 15(2): 734 - 750.
[Abstract] [Full Text] [PDF]

Home page
 Genes Dev.Home page
N. M. Hollingsworth and S. J. Brill
The Mus81 solution to resolution: generating meiotic crossovers without Holliday junctions
Genes & Dev., January 15, 2004; 18(2): 117 - 125.
[Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
J. L. Callahan, K. J. Andrews, V. A. Zakian, and C. H. Freudenreich
Mutations in Yeast Replication Proteins That Increase CAG/CTG Expansions Also Increase Repeat Fragility
Mol. Cell. Biol., November 1, 2003; 23(21): 7849 - 7860.
[Abstract] [Full Text] [PDF]

Home page
 Genes Dev.Home page
W. M. Fricke and S. J. Brill
Slx1--Slx4 is a second structure-specific endonuclease functionally redundant with Sgs1--Top3
Genes & Dev., July 15, 2003; 17(14): 1768 - 1778.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Sharma, J. A. Sommers, H. C. Driscoll, L. Uzdilla, T. M. Wilson, and R. M. Brosh Jr.
The Exonucleolytic and Endonucleolytic Cleavage Activities of Human Exonuclease 1 Are Stimulated by an Interaction with the Carboxyl-terminal Region of the Werner Syndrome Protein
J. Biol. Chem., June 20, 2003; 278(26): 23487 - 23496.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
T. de los Santos, N. Hunter, C. Lee, B. Larkin, J. Loidl, and N. M. Hollingsworth
The Mus81/Mms4 Endonuclease Acts Independently of Double-Holliday Junction Resolution to Promote a Distinct Subset of Crossovers During Meiosis in Budding Yeast
Genetics, May 1, 2003; 164(1): 81 - 94.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Liu and R. A. Bambara
Analysis of Human Flap Endonuclease 1 Mutants Reveals a Mechanism to Prevent Triplet Repeat Expansion
J. Biol. Chem., April 11, 2003; 278(16): 13728 - 13739.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Ayyagari, X. V. Gomes, D. A. Gordenin, and P. M. J. Burgers
Okazaki Fragment Maturation in Yeast. I. DISTRIBUTION OF FUNCTIONS BETWEEN FEN1 AND DNA2
J. Biol. Chem., January 10, 2003; 278(3): 1618 - 1625.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
A. Sundararajan, B.-S. Lee, and D. J. Garfinkel
The Rad27 (Fen-1) Nuclease Inhibits Ty1 Mobility in Saccharomyces cerevisiae
Genetics, January 1, 2003; 163(1): 55 - 67.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Lyonnais, C. Hounsou, M.-P. Teulade-Fichou, J. Jeusset, E. L. Cam, and G. Mirambeau
G-quartets assembly within a G-rich DNA flap. A possible event at the center of the HIV-1 genome
Nucleic Acids Res., December 1, 2002; 30(23): 5276 - 5283.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. A. Ranalli, S. Tom, and R. A. Bambara
AP Endonuclease 1 Coordinates Flap Endonuclease 1 and DNA Ligase I Activity in Long Patch Base Excision Repair
J. Biol. Chem., October 25, 2002; 277(44): 41715 - 41724.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.