Study of the location and function of N/sup 6/-methyladenosine in Rous sarcoma virus genomic RNA
N/sup 6/-methyladenosine (m/sup 6/A) residues are present as internal base modifications in most higher eukaryotic mRNAs; however, the biological function of this modification is not known. A method is described for localizing and quantitating m/sup 6/A within a large RNA molecule, the genomic RNA of Rous sarcoma virus (RSV). Specific fragments of /sup 32/P-labeled RSV RNA were isolated by hybridizations with complementary DNA restriction fragments spanning nucleotides 6185 and 8050. RNA was digested with RNAse and fingerprinted, and individual oligonucleotides were analyzed for the presence of m/sup 6/A by thin layer chromatography. With this technique, seven sites of methylation in this region of the RSV genome were localized at nucleotides 6394, 6447, 6507, 6718, 7414, 7424, and 8014. Further, m/sup 6/A was observed at two additional sites whose nucleotide assignments remain ambiguous. A clustering of two or more m/sup 6/A residues was seen at three positions within the RNA analyzed. Modification at certain sites was found to be heterogeneous, in that different molecules of RNA appeared to be methylated differently. An approach for directly studying the function of m/sup 6/A in RNA is presented.
- Research Organization:
- Johns Hopkins Univ., Baltimore, MD (USA)
- OSTI ID:
- 5349181
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ADENOSINE
BIOLOGICAL FUNCTIONS
BIOLOGICAL LOCALIZATION
RNA
METHYLATION
DNA
HYBRIDIZATION
NUCLEOTIDES
PHOSPHORUS 32
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
VIRUSES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
FUNCTIONS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEIC ACIDS
NUCLEOSIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RIBOSIDES
SEPARATION PROCESSES
550201* - Biochemistry- Tracer Techniques