Isolation and preliminary characterization of u. v. -sensitive mutants from the human cell line EUE

Journal Article · Sat Jan 01 04:00:00 EST 1983 · Carcinogenesis (N.Y.); (United States)

DOI:https://doi.org/10.1093/carcin/4.1.39· OSTI ID:5414928

Fiorio, R; Frosina, G; Abbondandolo, A

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), Journal Name: Carcinogenesis (N.Y.); (United States) Vol. 4:1; ISSN CRNGD
• Country of Publication: United States
• Language: English

Related Subjects

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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMALS
ANTIMETABOLITES
AUTORADIOGRAPHY
AZINES
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BROMOURACILS
BUDR
CELL CULTURES
CLONE CELLS
CYTOLOGICAL TECHNIQUES
DNA REPAIR
DRUGS
ELECTROMAGNETIC RADIATION
EMS
ESTERS
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MAN
MUTAGENS
MUTANTS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOTOSENSITIVITY
PRIMATES
PYRIMIDINES
RADIATION EFFECTS
RADIATIONS
RECOVERY
REPAIR
RIBOSIDES
SENSITIVITY
STRAND BREAKS
SULFONIC ACID ESTERS
THYMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ULTRAVIOLET RADIATION
URACILS
VERTEBRATES