Isolation and preliminary characterization of u. v. -sensitive mutants from the human cell line EUE
Five u.v. light-sensitive clones were isolated in the EUE cell line by means of a modified form of the original 5-bromodeoxyuridine (BUdR)-light method worked out by Puck and Kao for the isolation of nutritional mutants. A cell population was mutagenized with ethylmethanesulfonate. After the expression time, cells were u.v.-irradiated and incubated with BUdR to label excision patches in repair proficient cells. A subsequent irradiation with black light caused DNA strand breakage in BUdR-substituted cells. During BUdR treatment, hydroxyurea and a fluorochrome (Hoechst 33258) were added to possibly enhance the analogue incorporation into DNA and to increase the photolability of BUdR containing sequences, respectively. Out of 192 colonies selected with this method, 38 were isolated and tested for their u.v.-sensitivity. Five of them showed significant, reproducible differences with respect to the parental line. As a partial characterization, the five u.v.-sensitive clones were assayed for unscheduled (/sup 3/H)thymidine incorporation after exposure to u.v. light, by means of liquid scintillation spectrometry and autoradiography. In all clones. DNA repair synthesis was significantly decreased with respect to the parental line.
- Research Organization:
- Istituto di Mutagenesi e Differenziamento, Pisa, Italy
- OSTI ID:
- 5414928
- Journal Information:
- Carcinogenesis (N.Y.); (United States), Vol. 4:1
- Country of Publication:
- United States
- Language:
- English
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CLONE CELLS
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GENETIC EFFECTS
AUTORADIOGRAPHY
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CYTOLOGICAL TECHNIQUES
DNA REPAIR
MAN
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STRAND BREAKS
THYMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
ANIMALS
ANTIMETABOLITES
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BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
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CELL CULTURES
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ELECTROMAGNETIC RADIATION
ESTERS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MUTAGENS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PRIMATES
PYRIMIDINES
RADIATION EFFECTS
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SULFONIC ACID ESTERS
URACILS
VERTEBRATES
560301* - Chemicals Metabolism & Toxicology- Cells- (-1987)
560121 - Radiation Effects on Cells- External Source- (-1987)