Autoradiographic assay of mutants resistant to diphtheria toxin in mammalian cells in vitro

Journal Article · Mon Oct 01 00:00:00 EDT 1984 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.81.19.6124· OSTI ID:6135252

Ronen, A; Gingerich, J D; Duncan, A M.V.; Heddle, J A

Diptheria toxin kills mammalian cells by ribosylating elongation factor 2, a protein factor necessary for protein synthesis. The frequency of cells able to form colonies in the presence of the toxin can be used as an assay for mutation to diphtheria toxin resistance. Resistance to diphtheria toxin can also be detected autoradiographically in cells exposed to (/sup 3/H)leucine after treatment with the toxin. In cultures of Chinese hamster ovary cells, the frequency of such resistant cells is increased by exposure of the cells to ..gamma..-rays, ultraviolet light, ethylnitrosourea, mitomycin c, ethidium bromide, and 5-bromo-2'-deoxyuridine in a dose- and time-dependent manner. The resistant cells form discrete microcolonies if they are allowed to divide several times before intoxication which indicates that they are genuine mutants. The assay is potentially adaptable to any cell population that can be intoxicated with diphtheria toxin and labeled with (/sup 3/H)leucine, whether or not the cells can form colonies. It may be useful, therefore, for measuring mutation rates in slowly growing or nondividing cell populations such as breast, brain, and liver, as well as in cells that do divide but cannot be readily cloned, such as the colonic epithelium. 23 references, 6 figures.

• Research Organization: Ludwig Inst. for Cancer Research, Toronto, Ontario
• OSTI ID: 6135252
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 81:19
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
59 BASIC BIOLOGICAL SCIENCES
CHO CELLS
MUTATIONS
DEOXYURIDINE
BIOLOGICAL EFFECTS
DIPHTHERIA
TOXICITY
DISEASE RESISTANCE
MITOMYCIN
MUTANTS
AUTORADIOGRAPHY
NITROSOUREAS
DOSE-RESPONSE RELATIONSHIPS
IN VITRO
LEUCINE
TOXINS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
X RADIATION
AMINO ACIDS
ANIMAL CELLS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIGENS
ANTIMETABOLITES
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
AZINES
BACTERIAL DISEASES
CARBOXYLIC ACIDS
DISEASES
DRUGS
ELECTROMAGNETIC RADIATION
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
INFECTIOUS DISEASES
IONIZING RADIATIONS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MATERIALS
NITROSO COMPOUNDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RADIATIONS
RIBOSIDES
TOXIC MATERIALS
URACILS
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550701 - Microbiology- Tracer Techniques