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

Journal Article · Mon Oct 01 04:00:00 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), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 81:19; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550701 -- Microbiology-- Tracer Techniques
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACIDS
ANIMAL CELLS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIGENS
ANTIMETABOLITES
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
AUTORADIOGRAPHY
AZINES
BACTERIAL DISEASES
BIOLOGICAL EFFECTS
CARBOXYLIC ACIDS
CHO CELLS
DEOXYURIDINE
DIPHTHERIA
DISEASE RESISTANCE
DISEASES
DOSE-RESPONSE RELATIONSHIPS
DRUGS
ELECTROMAGNETIC RADIATION
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
IN VITRO
INFECTIOUS DISEASES
IONIZING RADIATIONS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LEUCINE
MATERIALS
MITOMYCIN
MUTANTS
MUTATIONS
NITROSO COMPOUNDS
NITROSOUREAS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RADIATIONS
RIBOSIDES
TOXIC MATERIALS
TOXICITY
TOXINS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
URACILS
X RADIATION