Demonstration of normal and mutant protein M1 subunits of deoxyGTP-resistant ribonucleotide reductase from mutant mouse lymphoma cells. Academic Article uri icon

Overview

abstract

  • From a mutagenized population of mouse T-lymphoma cells (S49) in continuous culture a cell line has been isolated (Ullman, B., Gudas, L. J., Clift, S. M., Martin, D. W., Jr. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 1074-1978) with ribonucleotide reductase activity that is inhibited only 50% by concentrations of dGTP which abolish wild type enzyme activity. Ribonucleotide reductase activity from this dGuo-L cell line retains its normal sensitivity to dATP. The partial sensitivity/partial resistance of the ribonucleotide reductase suggests that the dGuo-L cell line is heterozygous for ribonucleotide reductase, possessing one normal allele and one allele which codes for a dGTP-resistant enzyme. Both homologous and heterologous mixing experiments between the separated nonidentical subunits of ribonucleotide reductase, protein M1 and protein M2, from wild type and dGuo-L cells showed that the dGTP- feedback sensitivity was governed by the source of the protein M1. A partial resolution of two dGuo-L protein M1 components was achieved by chromatography on dextran blue-Sepharose. In order to resolve the two dGuo-L protein M1 components more completely, we introduced into dGuo-L cells a second mutation which conferred resistance of the ribonucleotide reductase to dATP, while the original dGTP resistance was maintained. The chromatography of protein M1 from this latter clone, dGuo-L-Aphid-G5, on dATP-Sepharose resolved two kinetically distinct protein M1 components. The first component was sensitive to dGTP inhibition but stimulated by dATP; the second was absolutely refractory to dGTP but sensitive to dATP inhibition. This confirms the hypothesis that the dGuo-L parent is heterozygous for protein M1, containing one wild type and one mutant allele.

publication date

  • October 10, 1981

Research

keywords

  • Deoxyguanine Nucleotides
  • Lymphoma
  • Mutation
  • Ribonucleotide Reductases

Identity

Scopus Document Identifier

  • 0019779607

PubMed ID

  • 7024266

Additional Document Info

volume

  • 256

issue

  • 19