Mismatch repair deficient human cells: Spontaneous and MNNG-induced mutational spectra in the HPRT gene

Aoy Tomita-Mitchell, Alexandra G. Kat, Luisa A. Marcelino, Xiao Cheng Li-Sucholeiki, Jacklene Goodluck-Griffith, William G. Thilly

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

We have determined both the spontaneous and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced mutational spectra in the HPRT gene of human cells (MT1) defective in the mismatch repair gene hMSH6 (GTBP). Eight of nine exons and nine of sixteen intronic flanking sequences were scanned, encompassing >900 bp of the HPRT gene. Mutant hotspots were detected and separated by differences in their melting temperatures using constant denaturant capillary electrophoresis (CDCE) or denaturing gradient gel electrophoresis (DGGE).A key finding of this work is that a high proportion of all HPRT inactivating mutations is represented by a small number of hotspots distributed over the exons and mRNA splice sites. Thirteen spontaneous hotspots and sixteen MNNG-induced hotspots accounted for 55% and 48% of all 6TG(R) point mutations, respectively. MNNG-induced hotspots were predominantly G:C→A:T transitions. The spontaneous spectrum of cells deficient in hMSH6 contained transversions (A:T→T:A, G:C→T:A, A:T→C:G), transitions (A:T→G:C), a plus-one insertion, and a minus-one deletion. Curiously, G:C→A:T transitions, which dominate human germinal and somatic point mutations were absent from the spontaneous hMSH6 spectra. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)125-138
Number of pages14
JournalMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume450
Issue number1-2
DOIs
StatePublished - May 30 2000

Keywords

  • Alkylation
  • CDCE
  • DGGE
  • High-fidelity PCR
  • Methylation
  • Mismatch repair
  • Mutational spectrum
  • hMSH6

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

Fingerprint Dive into the research topics of 'Mismatch repair deficient human cells: Spontaneous and MNNG-induced mutational spectra in the HPRT gene'. Together they form a unique fingerprint.

  • Cite this