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Abstract
An analysis of the methylated constituents of L cell mRNA by a combination of chromatographic methods and enzymatic treatments indicates that they comprise both 2′-0-methyl nucleosides and N 6-methyl adenine, and/or 1-methyl adenine, and suggests that the 2′-0-methyl nucleotides, Ym, are part of an unusual class of sequences forming the 5′ terminus of mRNA. These sequences seem to contain two 2′-0-methyl residues and a terminal residue that is not phosphorylated but, nevertheless, is blocked with respect to polynucleotide kinase reactivity. A strong candidate is a sequence of the type XppY 1mpY 2mpZp …, where X represents a blocking group which is itself occasionally methylated. The sequences isolated from total poly(A) + mRNA contain all four species of 2′-0-methylated nucleoside, indicating some variability among different mRNA species. The methylated sequences do not appear to be enriched in the mRNA which hybridizes with repetitive DNA. The average L cell mRNA molecule also contains three residues of N 6-methyl adenine. These residues are not part of the poly(A) segment, but appear to be located internal to the poly(A) near the 3′ end of the mRNA molecules.
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