The Zinc Linchpin Motif in the DNA Repair Glycosylase MUTYH: Identifying the Zn2+ Ligands and Roles in Damage Recognition and Repair

Nicole N. Nuñez, Cindy Khuu, C. Satheesan Babu, Steve J. Bertolani, Anisha N. Rajavel, Jensen E. Spear, Jeremy A. Armas, Jon D. Wright, Justin Siegel, Carmay Lim, Sheila S. David

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The DNA base excision repair (BER) glycosylase MUTYH prevents DNA mutations by catalyzing adenine (A) excision from inappropriately formed 8-oxoguanine (8-oxoG):A mismatches. The importance of this mutation suppression activity in tumor suppressor genes is underscored by the association of inherited variants of MUTYH with colorectal polyposis in a hereditary colorectal cancer syndrome known as MUTYH-associated polyposis, or MAP. Many of the MAP variants encompass amino acid changes that occur at positions surrounding the two-metal cofactor-binding sites of MUTYH. One of these cofactors, found in nearly all MUTYH orthologs, is a [4Fe-4S]2+ cluster coordinated by four Cys residues located in the N-terminal catalytic domain. We recently uncovered a second functionally relevant metal cofactor site present only in higher eukaryotic MUTYH orthologs: a Zn2+ ion coordinated by three Cys residues located within the extended interdomain connector (IDC) region of MUTYH that connects the N-terminal adenine excision and C-terminal 8-oxoG recognition domains. In this work, we identified a candidate for the fourth Zn2+ coordinating ligand using a combination of bioinformatics and computational modeling. In addition, using in vitro enzyme activity assays, fluorescence polarization DNA binding assays, circular dichroism spectroscopy, and cell-based rifampicin resistance assays, the functional impact of reduced Zn2+ chelation was evaluated. Taken together, these results illustrate the critical role that the "Zn2+ linchpin motif" plays in MUTYH repair activity by providing for proper engagement of the functional domains on the 8-oxoG:A mismatch required for base excision catalysis. The functional importance of the Zn2+ linchpin also suggests that adjacent MAP variants or exposure to environmental chemicals may compromise Zn2+ coordination, and ability of MUTYH to prevent disease.

Original languageEnglish (US)
Pages (from-to)13260-13271
Number of pages12
JournalJournal of the American Chemical Society
Volume140
Issue number41
DOIs
StatePublished - Oct 17 2018

Fingerprint

DNA Glycosylases
DNA Repair
Zinc
Assays
DNA
Repair
Ligands
Adenine
Hereditary Neoplastic Syndromes
Metals
Circular dichroism spectroscopy
Mutation
Fluorescence Polarization
Environmental Exposure
Enzyme Assays
Enzyme activity
Binding sites
Rifampin
Bioinformatics
Circular Dichroism

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

The Zinc Linchpin Motif in the DNA Repair Glycosylase MUTYH : Identifying the Zn2+ Ligands and Roles in Damage Recognition and Repair. / Nuñez, Nicole N.; Khuu, Cindy; Babu, C. Satheesan; Bertolani, Steve J.; Rajavel, Anisha N.; Spear, Jensen E.; Armas, Jeremy A.; Wright, Jon D.; Siegel, Justin; Lim, Carmay; David, Sheila S.

In: Journal of the American Chemical Society, Vol. 140, No. 41, 17.10.2018, p. 13260-13271.

Research output: Contribution to journalArticle

Nuñez, NN, Khuu, C, Babu, CS, Bertolani, SJ, Rajavel, AN, Spear, JE, Armas, JA, Wright, JD, Siegel, J, Lim, C & David, SS 2018, 'The Zinc Linchpin Motif in the DNA Repair Glycosylase MUTYH: Identifying the Zn2+ Ligands and Roles in Damage Recognition and Repair', Journal of the American Chemical Society, vol. 140, no. 41, pp. 13260-13271. https://doi.org/10.1021/jacs.8b06923
Nuñez, Nicole N. ; Khuu, Cindy ; Babu, C. Satheesan ; Bertolani, Steve J. ; Rajavel, Anisha N. ; Spear, Jensen E. ; Armas, Jeremy A. ; Wright, Jon D. ; Siegel, Justin ; Lim, Carmay ; David, Sheila S. / The Zinc Linchpin Motif in the DNA Repair Glycosylase MUTYH : Identifying the Zn2+ Ligands and Roles in Damage Recognition and Repair. In: Journal of the American Chemical Society. 2018 ; Vol. 140, No. 41. pp. 13260-13271.
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