The functional significance of various methylated nucleosides present in the nucleic acids of tumor cells and of DNA- and RNA-tumor virsues are being investigated by various immunochemical techniques. This approach employs antibodies specific for various methylated nucleosides which include 7-methylguanosine (m7G),N2,N2-dimethylguanosine (m2G), N6-methyladenosine (m6A), and 5-methylcytidine (m5C). Upon production, characterization, and purification, antibodies will be employed to isolate and characterize nucleic acid possessing the corresponding methylated nucleosides. Nucleic acids of particular interest include Adenovirus DNA and mRNAs, Rous Sarcoma Virus RNA and various bateriophages such as fd. The interaction between a specific anti-methylated nucleoside antibody and various nucleic acids provides unique immunochemical approaches for the (i) detection and (ii) quantitation of methylated constituents present in nucleic acids. Additionally, electron micrographs of nucleic acid:antibody complexes can be utilized (iii) to map the position of methylated nucleosides within a nucleic acid while immobilized antibodies can be employed (iv) to isolate hapten-containing (deoxy)oligonucleotides and nucleic acids. Last, the inhibition of specific enzyme activities associated with the metabolism of nucleic acids by the appropriate nucleic acid:antibody complexes will provide information regarding the (v) functional role(s) of the methylated nucleosides being examined.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA027801-07
Application #
3167833
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1982-09-01
Project End
1987-06-30
Budget Start
1986-03-01
Budget End
1987-06-30
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Munns, T W; Freeman, S K; Liszewski, M K et al. (1987) Distribution and specificity of nucleotide-reactive autoantibodies in human SLE. J Immunol 139:393-9
Munns, T W; Liszewski, M K; Freeman, S K (1986) Naturally occurring masked antibodies in murine sera recognize a component of the mitotic spindle apparatus. Biochem Biophys Res Commun 136:145-50