Quantitative analysis of nucleic acid alkylation in vitro and in cell culture
Thesis/Dissertation
·
OSTI ID:5776204
Chemical modification of polycytidylic acid with methyl methane-sulfonate has been studied by /sup 13/C NMR of the intact polymer, /sup 1/H NMR of the polymer, and /sup 1/H NMR, high-pressure liquid chromatography, and secondary-ion mass spectrometry (SIMS) of the nucleosides following enzymatic degradation. Using synthesized (/sup 2/H/sub 3/-methyl) labeled methylated nucleosides as internal references, the comparison of the relative amounts of products formed when calf thymus DNA is reacted with the weak carcinogen, methyl methanesulfonate and the potent carcinogen, 1-methyl-1-nitrosourea is achieved. Extension of this methodology to in vivo systems requires the recovery of highly purified DNA from the cells for accurate total methylation determinations by /sup 14/C incorporation which is achieved here by enzymatic and hydroxyapatite purifications. Upon alkylation of V79 Chinese hamster cells with methyl methanesulfonate, again only one methyl product, 7-methyl-2'-deoxyguanosine is detected.
- Research Organization:
- Purdue Univ., Lafayette, IN (USA)
- OSTI ID:
- 5776204
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201 -- Biochemistry-- Tracer Techniques
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKYLATION
ANIMAL CELLS
ANIMALS
BODY
CALVES
CARBON 13
CARBON ISOTOPES
CARCINOGENESIS
CARCINOGENS
CATTLE
CELL CULTURES
CHEMICAL ANALYSIS
CHEMICAL REACTIONS
CHO CELLS
CHROMATOGRAPHY
DEUTERIUM
DNA
DOMESTIC ANIMALS
ESTERS
EVEN-ODD NUCLEI
HYDROGEN ISOTOPES
IN VITRO
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
LYMPHATIC SYSTEM
MAGNETIC RESONANCE
MAMMALS
MASS SPECTROSCOPY
METHYL METHANESULFONATE
METHYLATION
MUTAGENS
NITROSO COMPOUNDS
NITROSOUREAS
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PATHOGENESIS
QUANTITATIVE CHEMICAL ANALYSIS
RESONANCE
RIBOSIDES
RUMINANTS
SEPARATION PROCESSES
SPECTROSCOPY
STABLE ISOTOPES
SULFONIC ACID ESTERS
THYMUS
VERTEBRATES
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKYLATION
ANIMAL CELLS
ANIMALS
BODY
CALVES
CARBON 13
CARBON ISOTOPES
CARCINOGENESIS
CARCINOGENS
CATTLE
CELL CULTURES
CHEMICAL ANALYSIS
CHEMICAL REACTIONS
CHO CELLS
CHROMATOGRAPHY
DEUTERIUM
DNA
DOMESTIC ANIMALS
ESTERS
EVEN-ODD NUCLEI
HYDROGEN ISOTOPES
IN VITRO
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
LYMPHATIC SYSTEM
MAGNETIC RESONANCE
MAMMALS
MASS SPECTROSCOPY
METHYL METHANESULFONATE
METHYLATION
MUTAGENS
NITROSO COMPOUNDS
NITROSOUREAS
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PATHOGENESIS
QUANTITATIVE CHEMICAL ANALYSIS
RESONANCE
RIBOSIDES
RUMINANTS
SEPARATION PROCESSES
SPECTROSCOPY
STABLE ISOTOPES
SULFONIC ACID ESTERS
THYMUS
VERTEBRATES