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PMID: 15673720 Published · epublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Functional characterization of two human MutY homolog (hMYH) missense mutations (R227W and V232F) that lie within the putative hMSH6 binding domain and are associated with hMYH polyposis.

Nucleic acids research ·Vol. 33 ·No. 2 ·2005-00-00 ·Pages 597-604

Bai H, Jones S, Guan X, Wilson TM, Sampson JR, Cheadle JP, Lu AL

Abstract

The base excision repair DNA glycosylase MutY homolog (MYH) is responsible for removing adenines misincorporated into DNA opposite guanine or 7,8-dihydro-8-oxo-guanine (8-oxoG), thereby preventing G:C to T:A mutations. Biallelic germline mutations in the human MYH gene predispose individuals to multiple colorectal adenomas and carcinoma. We have recently demonstrated that hMYH interacts with the mismatch repair protein hMSH6, and that the hMSH2/hMSH6 (hMutSalpha) heterodimer stimulates hMYH activity. Here, we characterize the functional effect of two missense mutations (R227W and V232F) associated with hMYH polyposis that lie within, or adjacent to, the putative hMSH6 binding domain. Neither missense mutation affects the physical interaction between hMYH and hMSH6. However, hMYH(R227W) has a severe defect in A/8-oxoG binding and glycosylase activities, while hMYH(V232F) has reduced A/8-oxoG binding and glycosylase activities. The glycosylase activity of the V232F mutant can be partially stimulated by hMutSalpha but cannot be restored to the wild-type level. Both mutants also fail to complement mutY-deficiency in Escherichia coli. These data define the pathogenic mechanisms underlying two further hMYH polyposis-associated mutations.

MeSH Terms
Adenomatous Polyposis Coli/genetics Adult Aged Amino Acid Sequence Binding Sites DNA/metabolism DNA Glycosylases/chemistry,genetics,metabolism DNA-Binding Proteins/metabolism Genetic Complementation Test Humans Male Molecular Sequence Data MutS Homolog 2 Protein Mutation, Missense Protein Structure, Tertiary Proto-Oncogene Proteins/metabolism
Chemicals
DNA-Binding Proteins G-T mismatch-binding protein Proto-Oncogene Proteins DNA DNA Glycosylases mutY adenine glycosylase MSH2 protein, human MutS Homolog 2 Protein
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Bai Haibo
Department of Biochemistry and Molecular Biology, University of Maryland Baltimore, MD, USA.
Jones Siân
Guan Xin
Wilson Teresa M
Sampson Julian R
Cheadle Jeremy P
Lu A-Lien
References (45)
45 references, click to expand
  1. Accumulation of premutagenic DNA lesions in mice defective in removal of oxidative base damage.
    Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13300-5 PMID: 10557315
  2. MSH2 and MSH6 are required for removal of adenine misincorporated opposite 8-oxo-guanine in S. cerevisiae.
    Mol Cell. 1999 Sep;4(3):439-44 PMID: 10518225
  3. Database of mouse strains carrying targeted mutations in genes affecting cellular responses to DNA damage. Version 4.
    Mutat Res. 2000 May 31;459(4):243-74 PMID: 10844240
  4. Repair of A/G and A/8-oxoG mismatches by MutY adenine DNA glycosylase.
    Methods Mol Biol. 2000;152:3-16 PMID: 10957964
  5. Human homolog of the MutY repair protein (hMYH) physically interacts with proteins involved in long patch DNA base excision repair.
    J Biol Chem. 2001 Feb 23;276(8):5547-55 PMID: 11092888
  6. Deficient DNA mismatch repair: a common etiologic factor for colon cancer.
    Hum Mol Genet. 2001 Apr;10(7):735-40 PMID: 11257106
  7. Inherited variants of MYH associated with somatic G:C-->T:A mutations in colorectal tumors.
    Nat Genet. 2002 Feb;30(2):227-32 PMID: 11818965
  8. Activation of human MutS homologs by 8-oxo-guanine DNA damage.
    J Biol Chem. 2002 Mar 8;277(10):8260-6 PMID: 11756455
  9. Repair of oxidative DNA damage: mechanisms and functions.
    Cell Biochem Biophys. 2001;35(2):141-70 PMID: 11892789
  10. Human MutY homolog, a DNA glycosylase involved in base excision repair, physically and functionally interacts with mismatch repair proteins human MutS homolog 2/human MutS homolog 6.
    J Biol Chem. 2002 Mar 29;277(13):11135-42 PMID: 11801590
  11. The mammalian mismatch repair pathway removes DNA 8-oxodGMP incorporated from the oxidized dNTP pool.
    Curr Biol. 2002 Jun 4;12(11):912-8 PMID: 12062055
  12. Biallelic germline mutations in MYH predispose to multiple colorectal adenoma and somatic G:C-->T:A mutations.
    Hum Mol Genet. 2002 Nov 1;11(23):2961-7 PMID: 12393807
  13. Multiple colorectal adenomas, classic adenomatous polyposis, and germ-line mutations in MYH.
    N Engl J Med. 2003 Feb 27;348(9):791-9 PMID: 12606733
  14. Insight into the functional consequences of inherited variants of the hMYH adenine glycosylase associated with colorectal cancer: complementation assays with hMYH variants and pre-steady-state kinetics of the corresponding mutated E.coli enzymes.
    J Mol Biol. 2003 Mar 21;327(2):431-43 PMID: 12628248
  15. Characterization of an Escherichia coli mutant MutY with a cysteine to alanine mutation at the iron-sulfur cluster domain.
    Biochemistry. 2003 Apr 8;42(13):3742-50 PMID: 12667065
  16. Germline mutations but not somatic changes at the MYH locus contribute to the pathogenesis of unselected colorectal cancers.
    Am J Pathol. 2003 May;162(5):1545-8 PMID: 12707038
  17. Autosomal recessive colorectal adenomatous polyposis due to inherited mutations of MYH.
    Lancet. 2003 Jul 5;362(9377):39-41 PMID: 12853198
  18. Exposing the MYtH about base excision repair and human inherited disease.
    Hum Mol Genet. 2003 Oct 15;12 Spec No 2:R159-65 PMID: 12915454
  19. Inherited variants in MYH are unlikely to contribute to the risk of lung carcinoma.
    Hum Genet. 2004 Jan;114(2):207-10 PMID: 14579148
  20. Structural basis for removal of adenine mispaired with 8-oxoguanine by MutY adenine DNA glycosylase.
    Nature. 2004 Feb 12;427(6975):652-6 PMID: 14961129
  21. Comprehensive analysis of the contribution of germline MYH variation to early-onset colorectal cancer.
    Int J Cancer. 2004 Apr 20;109(4):554-8 PMID: 14991577
  22. Identification of critical residues required for the mutation avoidance function of human MutY (hMYH) and implications in colorectal cancer.
    Cancer Lett. 2004 Mar 8;205(1):89-95 PMID: 15036665
  23. Deficiencies in mouse Myh and Ogg1 result in tumor predisposition and G to T mutations in codon 12 of the K-ras oncogene in lung tumors.
    Cancer Res. 2004 May 1;64(9):3096-102 PMID: 15126346
  24. MUTYH prevents OGG1 or APEX1 from inappropriately processing its substrate or reaction product with its C-terminal domain.
    Nucleic Acids Res. 2004;32(10):3198-211 PMID: 15199168
  25. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  26. Some mismatch repair activities in Escherichia coli.
    Proc Natl Acad Sci U S A. 1988 Dec;85(24):9674-8 PMID: 2974159
  27. Mechanistic studies of ionizing radiation and oxidative mutagenesis: genetic effects of a single 8-hydroxyguanine (7-hydro-8-oxoguanine) residue inserted at a unique site in a viral genome.
    Biochemistry. 1990 Jul 31;29(30):7024-32 PMID: 2223758
  28. Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxodG.
    Nature. 1991 Jan 31;349(6308):431-4 PMID: 1992344
  29. Two nicking enzyme systems specific for mismatch-containing DNA in nuclear extracts from human cells.
    J Biol Chem. 1991 Apr 5;266(10):6480-4 PMID: 1848856
  30. Site-specific mutagenesis using a gapped duplex vector: a study of translesion synthesis past 8-oxodeoxyguanosine in E. coli.
    Mutat Res. 1991 May;254(3):281-8 PMID: 2052015
  31. 8-Hydroxyguanine, an abundant form of oxidative DNA damage, causes G----T and A----C substitutions.
    J Biol Chem. 1992 Jan 5;267(1):166-72 PMID: 1730583
  32. A repair system for 8-oxo-7,8-dihydrodeoxyguanine.
    Biochemistry. 1992 Nov 17;31(45):10964-8 PMID: 1445834
  33. Single-stranded shuttle phagemid for mutagenesis studies in mammalian cells: 8-oxoguanine in DNA induces targeted G.C-->T.A transversions in simian kidney cells.
    Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1122-6 PMID: 8430083
  34. DNA determinants and substrate specificities of Escherichia coli MutY.
    J Biol Chem. 1995 Oct 6;270(40):23582-8 PMID: 7559523
  35. Mismatch repair in replication fidelity, genetic recombination, and cancer biology.
    Annu Rev Biochem. 1996;65:101-33 PMID: 8811176
  36. Cloning and characterization of hOGG1, a human homolog of the OGG1 gene of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8010-5 PMID: 9223305
  37. Molecular cloning and functional expression of a human cDNA encoding the antimutator enzyme 8-hydroxyguanine-DNA glycosylase.
    Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8016-20 PMID: 9223306
  38. 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) DNA glycosylase and AP lyase activities of hOGG1 protein and their substrate specificity.
    Mutat Res. 1997 Oct;385(1):75-82 PMID: 9372850
  39. Mouse embryonic stem cells carrying one or two defective Msh2 alleles respond abnormally to oxidative stress inflicted by low-level radiation.
    Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11915-20 PMID: 9751765
  40. MutY catalytic core, mutant and bound adenine structures define specificity for DNA repair enzyme superfamily.
    Nat Struct Biol. 1998 Dec;5(12):1058-64 PMID: 9846876
  41. Eukaryotic DNA mismatch repair.
    Curr Opin Genet Dev. 1999 Feb;9(1):89-96 PMID: 10072354
  42. Base excision repair of 8-hydroxyguanine protects DNA from endogenous oxidative stress.
    Biochimie. 1999 Jan-Feb;81(1-2):59-67 PMID: 10214911
  43. Differential subcellular localization of human MutY homolog (hMYH) and the functional activity of adenine:8-oxoguanine DNA glycosylase.
    Nucleic Acids Res. 1999 Sep 15;27(18):3638-44 PMID: 10471731
  44. Functional expression of hMYH, a human homolog of the Escherichia coli MutY protein.
    J Bacteriol. 1999 Oct;181(19):6210-3 PMID: 10498741
  45. Nuclear localization of the human mutY homologue hMYH.
    J Cell Biochem. 2000 Apr;77(4):666-77 PMID: 10771522
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gki209
Published
2005-00-00
Epub
2005-00-26
Pages
597-604
Language
English
Region
England
NLM ID
0411011
PMCID
PMC548354
Subset
IM
Grants
NCI NIH HHS · R01 CA095690 · United States
NCI NIH HHS · CA/ES78391 · United States
NCI NIH HHS · CA095690 · United States
External Links
PubMed source DOI: 10.1093/nar/gki209 PMC Full Text View journal details
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