Transfection of normal human and Chinese hamster DNA corrects diepoxybutane-induced chromosomal hypersensitivity of Fanconi anemia fibroblasts
Cultured cells from individuals affected with Fanconi anemia (FA) exhibit spontaneous chromosome breakage and hypersensitivity to the cell killing and clastogenic effects of the difunctional alkylating agent diepoxybutane (DEB). The authors report here the correction of both of these DEB-hypersensitivity phenotypes of FA cells achieved by cotransfection of normal placental of Chinese hamster lung cell DNA and the plasmid pSV2-neo-SVgpt. Transfectants were selected for clonogenic survival after treatment with DEB at a dose of 5 ..mu..gml. At this dose of DEB, the clonogenicity of normal fibroblasts was reduced to 50% and that of FA fibroblasts was reduced to zero. DEB-resistant (DEB/sup r/) colonies selected in this system exhibited a normal response to DEB-induced chromosome breakage and resistance to repeated DEB treatment. The neo and gpt sequences were detected by Southern blot analysis of DNA from one of four DEB/sup r/ colonies independently derived from transfection of human DNA and one of three DEB/sup r/ colonies independently derived from transfection of Chinese hamster DNA. The results demonstrate that DNA sequences that complement the two hallmark cellular phenotypes (cellular and chromosomal hypersensitivity to alkylating agents) of FA are present in human as well as Chinese hamster DNA. The cloning of these genes using transfection strategies can be expected to enable molecular characterization of FA
- Research Organization:
- Memorial Sloan-Kettering Cancer Center, New York, NY (USA)
- OSTI ID:
- 5173868
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:16
- Country of Publication:
- United States
- Language:
- English
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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKYLATING AGENTS
BIOLOGICAL EFFECTS
ANEMIAS
PATHOGENESIS
DNA
AUTORADIOGRAPHY
DNA SEQUENCING
CELL CULTURES
CELL KILLING
CHROMOSOMAL ABERRATIONS
ELECTROPHORESIS
FIBROBLASTS
HAMSTERS
HEREDITARY DISEASES
LUNGS
PATIENTS
PHOSPHORUS 32
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DISEASES
HEMIC DISEASES
ISOTOPES
LIGHT NUCLEI
MAMMALS
MUTATIONS
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RESPIRATORY SYSTEM
RODENTS
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
SYMPTOMS
VERTEBRATES
550401* - Genetics- Tracer Techniques
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