Comparative removal of pyrimidine dimers from human epidermal keratinocytes in vivo and in vitro

Journal Article · Sat Oct 01 04:00:00 EDT 1988 · J. Invest. Dermatol.; (United States)

DOI:https://doi.org/10.1111/1523-1747.ep12475699· OSTI ID:6541189

Reusch, M K; Meager, K; Leadon, S A; Hanawalt, P C

We have compared the kinetics for repair of UV-induced cyclobutane pyrimidine dimers in the DNA of keratinocytes in human skin and in cell culture. A small area of the buttocks of volunteers was exposed to UVB-irradiation and biopsies were taken at various time intervals. Epidermal keratinocytes in culture from the same subjects were exposed to UVC with doses chosen to elicit comparable yields of dimers in cellular DNA. The initial density of pyrimidine dimers and the kinetics of their removal were assessed utilizing the dimer-specific T4 endonuclease V and sedimentation of the unlabeled DNA through alkaline sucrose gradients. The position of DNA in the gradients was determined using a monoclonal antibody against random sequences of single-stranded DNA in a sensitive immunoassay. The initial dimer frequency was 3.9-6.7 per 10(8) daltons DNA. About 40% of the dimers were removed within 1 h, 70% in 6 h, and 90% in 24 h for both in vivo and in vitro samples. The early rapid removal phase may represent preferential repair of actively transcribed genes. Our findings reaffirm the usefulness and applicability of cell culture systems to model in vivo repair phenomena. The use of monoclonal antibodies to detect single-stranded DNA in alkaline sucrose gradients may be of value in a variety of studies involving human tissues in which it is not possible to use radioactive labeling of the DNA for the analysis.

• Research Organization: Stanford Univ., CA (USA)
• OSTI ID: 6541189
• Journal Information: J. Invest. Dermatol.; (United States), Journal Name: J. Invest. Dermatol.; (United States) Vol. 91:4; ISSN JIDEA
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BODY
COMPARATIVE EVALUATIONS
DNA
DNA REPAIR
ELECTROMAGNETIC RADIATION
ENZYME IMMUNOASSAY
EPIDERMIS
EPITHELIUM
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
IMMUNOASSAY
IN VITRO
IN VIVO
KERATIN
MAMMALS
MAN
MONOCLONAL ANTIBODIES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PRIMATES
PROTEINS
PYRIMIDINE DIMERS
RADIATION EFFECTS
RADIATIONS
RECOVERY
REMOVAL
REPAIR
SCLEROPROTEINS
SKIN
TISSUES
ULTRAVIOLET RADIATION
VERTEBRATES