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Lack of DNA enzymatic photoreactivation in HeLa cell-free extracts
- Source :
- FEBS Letters. (3):411-416
- Publisher :
- Published by Elsevier B.V.
-
Abstract
- Using a DNA-protein binding assay, we have previously identified and characterized a UV-damaged DNA recognition protein (UVDRP) from HeLa cells [(1991) Nucleic Acids Res. 19, 6413-6418]. In this report, the photoreactivating activity of UVDRP from the yeast, Saccharomyces cerevisiae, and HeLa cells was investigated. Although yeast and human cells are evolutionarily different from each other, both UVDRPs were conserved in the sense of their biochemical characteristics except that the yeast UVDRP also exhibited an enzymatic photoreactivating activity. A mammalian expression vector plasmid DNA carrying the bacterial chloramphenicol acetyltransferase (CAT) gene was UV irradiated in vitro followed immediately by exposure to photoreactivating light, and its transient expression in repair-deficient xeroderma pigmentosum (XP) cells was investigated. More than 80% of the CAT activity was inhibited by UV irradiation, which was partially restored (> 60%) by a partially purified yeast photolyase. In contrast, HeLa cell extracts did not express a photoreactivatable recovery from UV-induced inhibition of the CAT activity tested in the same system. This study has demonstrated the potential use of the DNA-mobility shift assay to investigate enzymatic photoreactivation, and has indicated the absence of the repair mechanism in human cells.
- Subjects :
- Xeroderma pigmentosum
Ultraviolet Rays
DNA repair
Saccharomyces cerevisiae
Biophysics
Photoreactivation
Biochemistry
Chromatography, Affinity
Cell Line
Fungal Proteins
HeLa
Chloramphenicol acetyltransferase
chemistry.chemical_compound
Structural Biology
Genetics
medicine
Humans
Damage-binding protein
DNA, Fungal
Photolyase
Molecular Biology
Cell Nucleus
Xeroderma Pigmentosum
Cell-Free System
biology
DNA, Neoplasm
Cell Biology
medicine.disease
biology.organism_classification
Molecular biology
Yeast
DNA-Binding Proteins
chemistry
Deoxyribodipyrimidine Photo-Lyase
DNA
DNA Damage
HeLa Cells
Subjects
Details
- Language :
- English
- ISSN :
- 00145793 and 64136418
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- FEBS Letters
- Accession number :
- edsair.doi.dedup.....a11a8998a9aeb451c9fbe9584fc2888c
- Full Text :
- https://doi.org/10.1016/0014-5793(93)80848-O