Ser 524 is a phosphorylation site in MUTYH and Ser 524 mutations alter 8-oxoguanine (OG): A mismatch recognition

Sucharita Kundu, Megan K. Brinkmeyer, Richard A. Eigenheer, Sheila S. David

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

MUTYH-associated polyposis (MAP) is a colorectal cancer predisposition syndrome that is caused by inherited biallelic mutations in the base excision repair (BER) gene, MUTYH. MUTYH is a DNA glycosylase that removes adenine (A) misinserted opposite 8-oxo-7,8-dihydro-2′-deoxyguanosine (OG). In this work, wild type (WT) MUTYH overexpressed using a baculovirus-driven insect cell expression system (BEVS) provided significantly higher levels of enzyme compared to bacterial overexpression. The isolated MUTYH enzyme was analyzed for potential post-translational modifications using mass spectrometry. An in vivo phosphorylation site was validated at Serine 524, which is located in the C-terminal OG recognition domain within the proliferating cell nuclear antigen (PCNA) binding region. Characterization of the phosphomimetic (S524D) and phosphoablating (S524A) mutants together with the observation that Ser 524 can be phosphorylated suggest that this residue may play an important regulatory role in vivo by altering stability and OG:A mismatch affinity.

Original languageEnglish (US)
Pages (from-to)1026-1037
Number of pages12
JournalDNA Repair
Volume9
Issue number10
DOIs
StatePublished - Oct 5 2010

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Keywords

  • 8-Oxoguanine
  • Adenine glycosylase
  • MAP
  • MUTYH
  • Phosphorylation
  • Post-translational modification

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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