Previous Article | Next Article 
Molecular and Cellular Biology, August 1999, p. 5373-5382, Vol. 19, No. 8
0270-7306/99/$04.00+0
A Novel Role in DNA Metabolism for the Binding
of Fen1/Rad27 to PCNA and Implications for Genetic Risk
Ronald
Gary,1,
Min S.
Park,1
John P.
Nolan,1
Helen L.
Cornelius,1
Olga G.
Kozyreva,2,
Hiep T.
Tran,2,§
Kirill S.
Lobachev,2
Michael A.
Resnick,2 and
Dmitry
A.
Gordenin2,*
Life Sciences Division, Los Alamos National
Laboratory, Los Alamos, New Mexico 87545,1
and Laboratory of Molecular Genetics, National Institute of
Environmental Health Sciences, Research Triangle Park, North
Carolina 277092
Received 5 March 1999/Returned for modification 21 April
1999/Accepted 13 May 1999
Fen1/Rad27 nuclease activity, which is important in DNA metabolism,
is stimulated by proliferating cell nuclear antigen (PCNA) in vitro.
The in vivo role of the PCNA interaction was investigated in the yeast
Rad27. A nuclease-defective rad27 mutation had a dominant-negative effect that was suppressed by a mutation in the PCNA
binding site, thereby demonstrating the importance of the Rad27-PCNA
interaction. The PCNA-binding defect alone had little effect on
mutation, recombination, and the methyl methanesulfonate (MMS) response
in repair-competent cells, but it greatly amplified the MMS sensitivity
of a rad51 mutant. Furthermore, the PCNA binding mutation
resulted in lethality when combined with a homozygous or even a
heterozygous pol3-01 mutation in the 3'
5' exonuclease domain of DNA polymerase
. These results suggest that phenotypically mild polymorphisms in DNA metabolic proteins can have dramatic consequences when combined.
*
Corresponding author. Mailing address: National
Institute of Environmental Health Sciences (NIEHS), Mail Drop D3-01,
101 TW Alexander Dr., P.O. Box 12233, Research Triangle Park, NC 27709. Phone: (919) 541-5190. Fax: (919) 541-7593. E-mail:
gordenin{at}niehs.nih.gov.

Present address: Department of Chemistry, University of Nevada, Las
Vegas, NV
89154.

Present address: Department of Biology, University of North
Carolina at Chapel Hill, Chapel Hill, NC 27599-3280.
§
Present address: LifeSensors, Inc., Malvern, PA
19355.
Molecular and Cellular Biology, August 1999, p. 5373-5382, Vol. 19, No. 8
0270-7306/99/$04.00+0
This article has been cited by other articles:
-
Karanja, K. K., Livingston, D. M.
(2009). C-Terminal Flap Endonuclease (rad27) Mutations: Lethal Interactions With a DNA Ligase I Mutation (cdc9-p) and Suppression by Proliferating Cell Nuclear Antigen (POL30) in Saccharomyces cerevisiae. Genetics
183: 63-78
[Abstract]
[Full Text]
-
Ma, W., Panduri, V., Sterling, J. F., Van Houten, B., Gordenin, D. A., Resnick, M. A.
(2009). The Transition of Closely Opposed Lesions to Double-Strand Breaks during Long-Patch Base Excision Repair Is Prevented by the Coordinated Action of DNA Polymerase {delta} and Rad27/Fen1. Mol. Cell. Biol.
29: 1212-1221
[Abstract]
[Full Text]
-
Xu, H., Boone, C., Brown, G. W.
(2007). Genetic Dissection of Parallel Sister-Chromatid Cohesion Pathways. Genetics
176: 1417-1429
[Abstract]
[Full Text]
-
Zheng, L., Dai, H., Qiu, J., Huang, Q., Shen, B.
(2007). Disruption of the FEN-1/PCNA Interaction Results in DNA Replication Defects, Pulmonary Hypoplasia, Pancytopenia, and Newborn Lethality in Mice. Mol. Cell. Biol.
27: 3176-3186
[Abstract]
[Full Text]
-
McNeill, D. R., Wilson, D. M. III
(2007). A Dominant-Negative Form of the Major Human Abasic Endonuclease Enhances Cellular Sensitivity to Laboratory and Clinical DNA-Damaging Agents. Mol Cancer Res
5: 61-70
[Abstract]
[Full Text]
-
Rossi, M. L., Bambara, R. A.
(2006). Reconstituted Okazaki Fragment Processing Indicates Two Pathways of Primer Removal. J. Biol. Chem.
281: 26051-26061
[Abstract]
[Full Text]
-
Refsland, E. W., Livingston, D. M.
(2005). Interactions Among DNA Ligase I, the Flap Endonuclease and Proliferating Cell Nuclear Antigen in the Expansion and Contraction of CAG Repeat Tracts in Yeast. Genetics
171: 923-934
[Abstract]
[Full Text]
-
Wells, R. D., Dere, R., Hebert, M. L., Napierala, M., Son, L. S.
(2005). Advances in mechanisms of genetic instability related to hereditary neurological diseases. Nucleic Acids Res
33: 3785-3798
[Abstract]
[Full Text]
-
Jin, Y. H., Garg, P., Stith, C. M. W., Al-Refai, H., Sterling, J. F., Murray, L. J. W., Kunkel, T. A., Resnick, M. A., Burgers, P. M., Gordenin, D. A.
(2005). The Multiple Biological Roles of the 3'->5' Exonuclease of Saccharomyces cerevisiae DNA Polymerase {delta} Require Switching between the Polymerase and Exonuclease Domains. Mol. Cell. Biol.
25: 461-471
[Abstract]
[Full Text]
-
Sharma, S., Sommers, J. A., Brosh, R. M. Jr
(2004). In vivo function of the conserved non-catalytic domain of Werner syndrome helicase in DNA replication. Hum Mol Genet
13: 2247-2261
[Abstract]
[Full Text]
-
Karthikeyan, G., Santos, J. H., Graziewicz, M. A., Copeland, W. C., Isaya, G., Houten, B. V., Resnick, M. A.
(2003). Reduction in frataxin causes progressive accumulation of mitochondrial damage. Hum Mol Genet
12: 3331-3342
[Abstract]
[Full Text]
-
Callahan, J. L., Andrews, K. J., Zakian, V. A., Freudenreich, C. H.
(2003). Mutations in Yeast Replication Proteins That Increase CAG/CTG Expansions Also Increase Repeat Fragility. Mol. Cell. Biol.
23: 7849-7860
[Abstract]
[Full Text]
-
Spiro, C., McMurray, C. T.
(2003). Nuclease-Deficient FEN-1 Blocks Rad51/BRCA1-Mediated Repair and Causes Trinucleotide Repeat Instability. Mol. Cell. Biol.
23: 6063-6074
[Abstract]
[Full Text]
-
Galli, A., Cervelli, T., Schiestl, R. H.
(2003). Characterization of the Hyperrecombination Phenotype of the pol3-t Mutation of Saccharomyces cerevisiae. Genetics
164: 65-79
[Abstract]
[Full Text]
-
Liu, Y., Bambara, R. A.
(2003). Analysis of Human Flap Endonuclease 1 Mutants Reveals a Mechanism to Prevent Triplet Repeat Expansion. J. Biol. Chem.
278: 13728-13739
[Abstract]
[Full Text]
-
Ayyagari, R., Gomes, X. V., Gordenin, D. A., Burgers, P. M. J.
(2003). Okazaki Fragment Maturation in Yeast. I. DISTRIBUTION OF FUNCTIONS BETWEEN FEN1 AND DNA2. J. Biol. Chem.
278: 1618-1625
[Abstract]
[Full Text]
-
Jin, Y. H., Ayyagari, R., Resnick, M. A., Gordenin, D. A., Burgers, P. M. J.
(2003). Okazaki Fragment Maturation in Yeast. II. COOPERATION BETWEEN THE POLYMERASE AND 3'-5'-EXONUCLEASE ACTIVITIES OF POL delta IN THE CREATION OF A LIGATABLE NICK. J. Biol. Chem.
278: 1626-1633
[Abstract]
[Full Text]
-
Karthikeyan, G., Lewis, L. K., Resnick, M. A.
(2002). The mitochondrial protein frataxin prevents nuclear damage. Hum Mol Genet
11: 1351-1362
[Abstract]
[Full Text]
-
Shibata, Y., Nakamura, T.
(2002). Defective Flap Endonuclease 1 Activity in Mammalian Cells Is Associated with Impaired DNA Repair and Prolonged S Phase Delay. J. Biol. Chem.
277: 746-754
[Abstract]
[Full Text]
-
Lewis, L. K., Karthikeyan, G., Westmoreland, J. W., Resnick, M. A.
(2002). Differential Suppression of DNA Repair Deficiencies of Yeast rad50, mre11 and xrs2 Mutants by EXO1 and TLC1 (the RNA Component of Telomerase). Genetics
160: 49-62
[Abstract]
[Full Text]
-
Young, S. M. Jr., Samulski, R. J.
(2001). Adeno-associated virus (AAV) site-specific recombination does not require a Rep-dependent origin of replication within the AAV terminal repeat. Proc. Natl. Acad. Sci. USA
10.1073/pnas.241508998v1
[Abstract]
[Full Text]
-
Xie, Y., Liu, Y., Argueso, J. L., Henricksen, L. A., Kao, H.-I, Bambara, R. A., Alani, E.
(2001). Identification of rad27 Mutations That Confer Differential Defects in Mutation Avoidance, Repeat Tract Instability, and Flap Cleavage. Mol. Cell. Biol.
21: 4889-4899
[Abstract]
[Full Text]
-
Debrauwere, H., Loeillet, S., Lin, W., Lopes, J., Nicolas, A.
(2001). Links between replication and recombination in Saccharomyces cerevisiae: A hypersensitive requirement for homologous recombination in the absence of Rad27 activity. Proc. Natl. Acad. Sci. USA
98: 8263-8269
[Abstract]
[Full Text]
-
Hanna, J. S., Kroll, E. S., Lundblad, V., Spencer, F. A.
(2001). Saccharomyces cerevisiae CTF18 and CTF4 Are Required for Sister Chromatid Cohesion. Mol. Cell. Biol.
21: 3144-3158
[Abstract]
[Full Text]
-
Jin, Y. H., Obert, R., Burgers, P. M. J., Kunkel, T. A., Resnick, M. A., Gordenin, D. A.
(2001). The 3'right-arrow5' exonuclease of DNA polymerase delta can substitute for the 5' flap endonuclease Rad27/Fen1 in processing Okazaki fragments and preventing genome instability. Proc. Natl. Acad. Sci. USA
10.1073/pnas.091095198v1
[Abstract]
[Full Text]
-
Kirchner, J. M., Tran, H., Resnick, M. A.
(2000). A DNA Polymerase {epsilon} Mutant That Specifically Causes +1 Frameshift Mutations Within Homonucleotide Runs in Yeast. Genetics
155: 1623-1632
[Abstract]
[Full Text]
-
Kang, H.-Y., Choi, E., Bae, S.-H., Lee, K.-H., Gim, B.-S., Kim, H.-D., Park, C., MacNeill, S. A., Seo, Y.-S.
(2000). Genetic Analyses of Schizosaccharomyces pombe dna2+ Reveal That Dna2 Plays an Essential Role in Okazaki Fragment Metabolism. Genetics
155: 1055-1067
[Abstract]
[Full Text]
-
Sokolsky, T., Alani, E.
(2000). EXO1 and MSH6 Are High-Copy Suppressors of Conditional Mutations in the MSH2 Mismatch Repair Gene of Saccharomyces cerevisiae. Genetics
155: 589-599
[Abstract]
[Full Text]
-
Tom, S., Henricksen, L. A., Bambara, R. A.
(2000). Mechanism Whereby Proliferating Cell Nuclear Antigen Stimulates Flap Endonuclease 1. J. Biol. Chem.
275: 10498-10505
[Abstract]
[Full Text]
-
Clark, A. B., Valle, F., Drotschmann, K., Gary, R. K., Kunkel, T. A.
(2000). Functional Interaction of Proliferating Cell Nuclear Antigen with MSH2-MSH6 and MSH2-MSH3 Complexes. J. Biol. Chem.
275: 36498-36501
[Abstract]
[Full Text]
-
Bae, S.-H., Seo, Y.-S.
(2000). Characterization of the Enzymatic Properties of the Yeast Dna2 Helicase/Endonuclease Suggests a New Model for Okazaki Fragment Processing. J. Biol. Chem.
275: 38022-38031
[Abstract]
[Full Text]
-
Qiu, J., Li, X., Frank, G., Shen, B.
(2001). Cell Cycle-dependent and DNA Damage-inducible Nuclear Localization of FEN-1 Nuclease Is Consistent with Its Dual Functions in DNA Replication and Repair. J. Biol. Chem.
276: 4901-4908
[Abstract]
[Full Text]
-
Parker, A., Gu, Y., Mahoney, W., Lee, S.-H., Singh, K. K., Lu, A-L.
(2001). Human Homolog of the MutY Repair Protein (hMYH) Physically Interacts with Proteins Involved in Long Patch DNA Base Excision Repair. J. Biol. Chem.
276: 5547-5555
[Abstract]
[Full Text]
-
Stucki, M., Jonsson, Z. O., Hubscher, U.
(2001). In Eukaryotic Flap Endonuclease 1, the C Terminus Is Essential for Substrate Binding. J. Biol. Chem.
276: 7843-7849
[Abstract]
[Full Text]
-
Frank, G., Qiu, J., Zheng, L., Shen, B.
(2001). Stimulation of Eukaryotic Flap Endonuclease-1 Activities by Proliferating Cell Nuclear Antigen (PCNA) Is Independent of Its in Vitro Interaction via a Consensus PCNA Binding Region. J. Biol. Chem.
276: 36295-36302
[Abstract]
[Full Text]
-
Jin, Y. H., Obert, R., Burgers, P. M. J., Kunkel, T. A., Resnick, M. A., Gordenin, D. A.
(2001). The 3'right-arrow5' exonuclease of DNA polymerase delta can substitute for the 5' flap endonuclease Rad27/Fen1 in processing Okazaki fragments and preventing genome instability. Proc. Natl. Acad. Sci. USA
98: 5122-5127
[Abstract]
[Full Text]
-
Young, S. M. Jr., Samulski, R. J.
(2001). Adeno-associated virus (AAV) site-specific recombination does not require a Rep-dependent origin of replication within the AAV terminal repeat. Proc. Natl. Acad. Sci. USA
98: 13525-13530
[Abstract]
[Full Text]