Distinct functional consequences of MUTYH variants associated with colorectal cancer

Damaged DNA affinity, glycosylase activity and interaction with PCNA and Hus1

Megan K. Brinkmeyer, Sheila S. David

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

MUTYH is a base excision repair (BER) enzyme that prevents mutations in DNA associated with 8-oxoguanine (OG) by catalyzing the removal of adenine from inappropriately formed OG:A base-pairs. Germline mutations in the MUTYH gene are linked to colorectal polyposis and a high risk of colorectal cancer, a syndrome referred to as MUTYH-associated polyposis (MAP). There are over 300 different MUTYH mutations associated with MAP and a large fraction of these gene changes code for missense MUTYH variants. Herein, the adenine glycosylase activity, mismatch recognition properties, and interaction with relevant protein partners of human MUTYH and five MAP variants (R295C, P281L, Q324H, P502L, and R520Q) were examined. P281L MUTYH was found to be severely compromised both in DNA binding and base excision activity, consistent with the location of this variation in the iron-sulfur cluster (FCL) DNA binding motif of MUTYH. Both R295C and R520Q MUTYH were found to have low fractions of active enzyme, compromised affinity for damaged DNA, and reduced rates for adenine excision. In contrast, both Q324H and P502L MUTYH function relatively similarly to WT MUTYH in both binding and glycosylase assays. However, P502L and R520Q exhibited reduced affinity for PCNA (proliferation cell nuclear antigen), consistent with their location in the PCNA-binding motif of MUTYH. Whereas, only Q324H, and not R295C, was found to have reduced affinity for Hus1 of the Rad9-Hus1-Rad1 complex, despite both being localized to the same region implicated for interaction with Hus1. These results underscore the diversity of functional consequences due to MUTYH variants that may impact the progression of MAP.

Original languageEnglish (US)
Pages (from-to)39-51
Number of pages13
JournalDNA Repair
Volume34
DOIs
StatePublished - Oct 1 2015

Fingerprint

DNA Glycosylases
Nuclear Antigens
Cell proliferation
Colorectal Neoplasms
Cell Proliferation
Adenine
DNA
Mutation
Nucleotide Motifs
Germ-Line Mutation
Genes
Enzymes
Sulfur
Base Pairing
DNA Repair
Iron
Assays
Repair
Proteins
8-hydroxyguanine

Keywords

  • 8-Oxoguanine
  • Base excision repair (BER)
  • Cancer variants
  • DNA glycosylase
  • Enzyme kinetics
  • Hus1
  • MUTYH-associated polyposis (MAP)
  • PCNA
  • Single nucleotide polymorphisms

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Distinct functional consequences of MUTYH variants associated with colorectal cancer : Damaged DNA affinity, glycosylase activity and interaction with PCNA and Hus1. / Brinkmeyer, Megan K.; David, Sheila S.

In: DNA Repair, Vol. 34, 01.10.2015, p. 39-51.

Research output: Contribution to journalArticle

@article{b42a992c393f4bcb9e4a21b0e152f7c6,
title = "Distinct functional consequences of MUTYH variants associated with colorectal cancer: Damaged DNA affinity, glycosylase activity and interaction with PCNA and Hus1",
abstract = "MUTYH is a base excision repair (BER) enzyme that prevents mutations in DNA associated with 8-oxoguanine (OG) by catalyzing the removal of adenine from inappropriately formed OG:A base-pairs. Germline mutations in the MUTYH gene are linked to colorectal polyposis and a high risk of colorectal cancer, a syndrome referred to as MUTYH-associated polyposis (MAP). There are over 300 different MUTYH mutations associated with MAP and a large fraction of these gene changes code for missense MUTYH variants. Herein, the adenine glycosylase activity, mismatch recognition properties, and interaction with relevant protein partners of human MUTYH and five MAP variants (R295C, P281L, Q324H, P502L, and R520Q) were examined. P281L MUTYH was found to be severely compromised both in DNA binding and base excision activity, consistent with the location of this variation in the iron-sulfur cluster (FCL) DNA binding motif of MUTYH. Both R295C and R520Q MUTYH were found to have low fractions of active enzyme, compromised affinity for damaged DNA, and reduced rates for adenine excision. In contrast, both Q324H and P502L MUTYH function relatively similarly to WT MUTYH in both binding and glycosylase assays. However, P502L and R520Q exhibited reduced affinity for PCNA (proliferation cell nuclear antigen), consistent with their location in the PCNA-binding motif of MUTYH. Whereas, only Q324H, and not R295C, was found to have reduced affinity for Hus1 of the Rad9-Hus1-Rad1 complex, despite both being localized to the same region implicated for interaction with Hus1. These results underscore the diversity of functional consequences due to MUTYH variants that may impact the progression of MAP.",
keywords = "8-Oxoguanine, Base excision repair (BER), Cancer variants, DNA glycosylase, Enzyme kinetics, Hus1, MUTYH-associated polyposis (MAP), PCNA, Single nucleotide polymorphisms",
author = "Brinkmeyer, {Megan K.} and David, {Sheila S.}",
year = "2015",
month = "10",
day = "1",
doi = "10.1016/j.dnarep.2015.08.001",
language = "English (US)",
volume = "34",
pages = "39--51",
journal = "DNA Repair",
issn = "1568-7864",
publisher = "Elsevier",

}

TY - JOUR

T1 - Distinct functional consequences of MUTYH variants associated with colorectal cancer

T2 - Damaged DNA affinity, glycosylase activity and interaction with PCNA and Hus1

AU - Brinkmeyer, Megan K.

AU - David, Sheila S.

PY - 2015/10/1

Y1 - 2015/10/1

N2 - MUTYH is a base excision repair (BER) enzyme that prevents mutations in DNA associated with 8-oxoguanine (OG) by catalyzing the removal of adenine from inappropriately formed OG:A base-pairs. Germline mutations in the MUTYH gene are linked to colorectal polyposis and a high risk of colorectal cancer, a syndrome referred to as MUTYH-associated polyposis (MAP). There are over 300 different MUTYH mutations associated with MAP and a large fraction of these gene changes code for missense MUTYH variants. Herein, the adenine glycosylase activity, mismatch recognition properties, and interaction with relevant protein partners of human MUTYH and five MAP variants (R295C, P281L, Q324H, P502L, and R520Q) were examined. P281L MUTYH was found to be severely compromised both in DNA binding and base excision activity, consistent with the location of this variation in the iron-sulfur cluster (FCL) DNA binding motif of MUTYH. Both R295C and R520Q MUTYH were found to have low fractions of active enzyme, compromised affinity for damaged DNA, and reduced rates for adenine excision. In contrast, both Q324H and P502L MUTYH function relatively similarly to WT MUTYH in both binding and glycosylase assays. However, P502L and R520Q exhibited reduced affinity for PCNA (proliferation cell nuclear antigen), consistent with their location in the PCNA-binding motif of MUTYH. Whereas, only Q324H, and not R295C, was found to have reduced affinity for Hus1 of the Rad9-Hus1-Rad1 complex, despite both being localized to the same region implicated for interaction with Hus1. These results underscore the diversity of functional consequences due to MUTYH variants that may impact the progression of MAP.

AB - MUTYH is a base excision repair (BER) enzyme that prevents mutations in DNA associated with 8-oxoguanine (OG) by catalyzing the removal of adenine from inappropriately formed OG:A base-pairs. Germline mutations in the MUTYH gene are linked to colorectal polyposis and a high risk of colorectal cancer, a syndrome referred to as MUTYH-associated polyposis (MAP). There are over 300 different MUTYH mutations associated with MAP and a large fraction of these gene changes code for missense MUTYH variants. Herein, the adenine glycosylase activity, mismatch recognition properties, and interaction with relevant protein partners of human MUTYH and five MAP variants (R295C, P281L, Q324H, P502L, and R520Q) were examined. P281L MUTYH was found to be severely compromised both in DNA binding and base excision activity, consistent with the location of this variation in the iron-sulfur cluster (FCL) DNA binding motif of MUTYH. Both R295C and R520Q MUTYH were found to have low fractions of active enzyme, compromised affinity for damaged DNA, and reduced rates for adenine excision. In contrast, both Q324H and P502L MUTYH function relatively similarly to WT MUTYH in both binding and glycosylase assays. However, P502L and R520Q exhibited reduced affinity for PCNA (proliferation cell nuclear antigen), consistent with their location in the PCNA-binding motif of MUTYH. Whereas, only Q324H, and not R295C, was found to have reduced affinity for Hus1 of the Rad9-Hus1-Rad1 complex, despite both being localized to the same region implicated for interaction with Hus1. These results underscore the diversity of functional consequences due to MUTYH variants that may impact the progression of MAP.

KW - 8-Oxoguanine

KW - Base excision repair (BER)

KW - Cancer variants

KW - DNA glycosylase

KW - Enzyme kinetics

KW - Hus1

KW - MUTYH-associated polyposis (MAP)

KW - PCNA

KW - Single nucleotide polymorphisms

UR - http://www.scopus.com/inward/record.url?scp=84942818696&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84942818696&partnerID=8YFLogxK

U2 - 10.1016/j.dnarep.2015.08.001

DO - 10.1016/j.dnarep.2015.08.001

M3 - Article

VL - 34

SP - 39

EP - 51

JO - DNA Repair

JF - DNA Repair

SN - 1568-7864

ER -