Uracil DNA N-glycosylase Promotes Assembly of Human Centromere Protein A

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2011 Mar 15

PMID 21399697

Citations 12

Authors

Samantha G Zeitlin

Brian R Chapados

Norman M Baker

Caroline Tai

Geir Slupphaug

Jean Y J Wang

Affiliations

Soon will be listed here.

Abstract

Uracil is removed from DNA by the conserved enzyme uracil DNA N-glycosylase (UNG). Previously, we observed that inhibiting UNG in Xenopus egg extracts blocked assembly of CENP-A, a histone H3 variant. CENP-A is an essential protein in all species, since it is required for chromosome segregation during mitosis. Thus, the implication of UNG in CENP-A assembly implies that UNG would also be essential, but UNG mutants lacking catalytic activity are viable in all species. In this paper, we present evidence that UNG2 colocalizes with CENP-A and H2AX phosphorylation at centromeres in normally cycling cells. Reduction of UNG2 in human cells blocks CENP-A assembly, and results in reduced cell proliferation, associated with increased frequencies of mitotic abnormalities and rapid cell death. Overexpression of UNG2 induces high levels of CENP-A assembly in human cells. Using a multiphoton laser approach, we demonstrate that UNG2 is rapidly recruited to sites of DNA damage. Taken together, our data are consistent with a model in which the N-terminus of UNG2 interacts with the active site of the enzyme and with chromatin.

Citing Articles

HIV-1 Vpr Functions in Primary CD4 T Cells.

Vanegas-Torres C, Schindler M Viruses. 2024; 16(3).

PMID: 38543785 PMC: 10975730. DOI: 10.3390/v16030420.

Histone variants: The unsung guardians of the genome.

Phillips E, Gunjan A DNA Repair (Amst). 2022; 112:103301.

PMID: 35220000 PMC: 9248346. DOI: 10.1016/j.dnarep.2022.103301.

Identification of a Chemotherapeutic Lead Molecule for the Potential Disruption of the FAM72A-UNG2 Interaction to Interfere with Genome Stability, Centromere Formation, and Genome Editing.

Renganathan S, Pramanik S, Ekambaram R, Kutzner A, Kim P, Heese K Cancers (Basel). 2021; 13(22).

PMID: 34831023 PMC: 8616359. DOI: 10.3390/cancers13225870.

RPA2 winged-helix domain facilitates UNG-mediated removal of uracil from ssDNA; implications for repair of mutagenic uracil at the replication fork.

Kavli B, Iveland T, Buchinger E, Hagen L, Liabakk N, Aas P Nucleic Acids Res. 2021; 49(7):3948-3966.

PMID: 33784377 PMC: 8053108. DOI: 10.1093/nar/gkab195.

Deoxyuracil in DNA and disease: Genomic signal or managed situation?.

Chon J, Field M, Stover P DNA Repair (Amst). 2019; 77:36-44.

PMID: 30875637 PMC: 6481626. DOI: 10.1016/j.dnarep.2019.02.014.

References

Slupphaug G, Eftedal I, Kavli B, Bharati S, Helle N, Haug T . Properties of a recombinant human uracil-DNA glycosylase from the UNG gene and evidence that UNG encodes the major uracil-DNA glycosylase. Biochemistry. 1995; 34(1):128-38. DOI: 10.1021/bi00001a016. View

Handa P, Acharya N, Varshney U . Effects of mutations at tyrosine 66 and asparagine 123 in the active site pocket of Escherichia coli uracil DNA glycosylase on uracil excision from synthetic DNA oligomers: evidence for the occurrence of long-range interactions between the enzyme.... Nucleic Acids Res. 2002; 30(14):3086-95. PMC: 135746. DOI: 10.1093/nar/gkf425. View

Nilsen H, Rosewell I, Robins P, Skjelbred C, Andersen S, Slupphaug G . Uracil-DNA glycosylase (UNG)-deficient mice reveal a primary role of the enzyme during DNA replication. Mol Cell. 2000; 5(6):1059-65. DOI: 10.1016/s1097-2765(00)80271-3. View

Otterlei M, Warbrick E, Nagelhus T, Haug T, Slupphaug G, Akbari M . Post-replicative base excision repair in replication foci. EMBO J. 1999; 18(13):3834-44. PMC: 1171460. DOI: 10.1093/emboj/18.13.3834. View

Begum N, Kinoshita K, Kakazu N, Muramatsu M, Nagaoka H, Shinkura R . Uracil DNA glycosylase activity is dispensable for immunoglobulin class switch. Science. 2004; 305(5687):1160-3. DOI: 10.1126/science.1098444. View

Fischer J, Muller-Weeks S, Caradonna S . Proteolytic degradation of the nuclear isoform of uracil-DNA glycosylase occurs during the S phase of the cell cycle. DNA Repair (Amst). 2004; 3(5):505-13. DOI: 10.1016/j.dnarep.2004.01.012. View

Pulukuri S, Knost J, Estes N, Rao J . Small interfering RNA-directed knockdown of uracil DNA glycosylase induces apoptosis and sensitizes human prostate cancer cells to genotoxic stress. Mol Cancer Res. 2009; 7(8):1285-93. PMC: 2730435. DOI: 10.1158/1541-7786.MCR-08-0508. View

Nakamura N, Morinaga H, Kikuchi M, Yonekura S, Ishii N, Yamamoto K . Cloning and characterization of uracil-DNA glycosylase and the biological consequences of the loss of its function in the nematode Caenorhabditis elegans. Mutagenesis. 2008; 23(5):407-13. DOI: 10.1093/mutage/gen030. View

Kruman I, Schwartz E, Kruman Y, Cutler R, Zhu X, Greig N . Suppression of uracil-DNA glycosylase induces neuronal apoptosis. J Biol Chem. 2004; 279(42):43952-60. DOI: 10.1074/jbc.M408025200. View

10.

Parikh S, Mol C, Slupphaug G, Bharati S, Krokan H, Tainer J . Base excision repair initiation revealed by crystal structures and binding kinetics of human uracil-DNA glycosylase with DNA. EMBO J. 1998; 17(17):5214-26. PMC: 1170849. DOI: 10.1093/emboj/17.17.5214. View

11.

An Q, Robins P, Lindahl T, Barnes D . C --> T mutagenesis and gamma-radiation sensitivity due to deficiency in the Smug1 and Ung DNA glycosylases. EMBO J. 2005; 24(12):2205-13. PMC: 1150883. DOI: 10.1038/sj.emboj.7600689. View

12.

Begum N, Izumi N, Nishikori M, Nagaoka H, Shinkura R, Honjo T . Requirement of non-canonical activity of uracil DNA glycosylase for class switch recombination. J Biol Chem. 2006; 282(1):731-42. DOI: 10.1074/jbc.M607439200. View

13.

Imai K, Slupphaug G, Lee W, Revy P, Nonoyama S, Catalan N . Human uracil-DNA glycosylase deficiency associated with profoundly impaired immunoglobulin class-switch recombination. Nat Immunol. 2003; 4(10):1023-8. DOI: 10.1038/ni974. View

14.

Dianov G, Timchenko T, Sinitsina O, Kuzminov A, Medvedev O, Salganik R . Repair of uracil residues closely spaced on the opposite strands of plasmid DNA results in double-strand break and deletion formation. Mol Gen Genet. 1991; 225(3):448-52. DOI: 10.1007/BF00261686. View

15.

Pan-Hammarstrom Q, Zhao Y, Hammarstrom L . Class switch recombination: a comparison between mouse and human. Adv Immunol. 2007; 93:1-61. DOI: 10.1016/S0065-2776(06)93001-6. View

16.

Andersen S, Ericsson M, Dai H, Pena-Diaz J, Slupphaug G, Nilsen H . Monoclonal B-cell hyperplasia and leukocyte imbalance precede development of B-cell malignancies in uracil-DNA glycosylase deficient mice. DNA Repair (Amst). 2005; 4(12):1432-41. DOI: 10.1016/j.dnarep.2005.08.004. View

17.

Studebaker A, Ariza M, Williams M . Depletion of uracil-DNA glycosylase activity is associated with decreased cell proliferation. Biochem Biophys Res Commun. 2005; 334(2):509-15. DOI: 10.1016/j.bbrc.2005.06.118. View

18.

Nilsen H, Otterlei M, Haug T, Solum K, Nagelhus T, Skorpen F . Nuclear and mitochondrial uracil-DNA glycosylases are generated by alternative splicing and transcription from different positions in the UNG gene. Nucleic Acids Res. 1997; 25(4):750-5. PMC: 146498. DOI: 10.1093/nar/25.4.750. View

19.

Zeitlin S, Baker N, Chapados B, Soutoglou E, Wang J, Berns M . Double-strand DNA breaks recruit the centromeric histone CENP-A. Proc Natl Acad Sci U S A. 2009; 106(37):15762-7. PMC: 2747192. DOI: 10.1073/pnas.0908233106. View

20.

Foltz D, Jansen L, Bailey A, Yates 3rd J, Bassett E, Wood S . Centromere-specific assembly of CENP-a nucleosomes is mediated by HJURP. Cell. 2009; 137(3):472-84. PMC: 2747366. DOI: 10.1016/j.cell.2009.02.039. View