Histone MRNAs Contain Blocked and Methylated 5' Terminal Sequences but Lack Methylated Nucleosides at Internal Positions

Histone mRNA, labeled with 32P or 3H-methionine during the S phase of partially synchronized HeLa cells, was isolated from the polyribosomes and purified as a "9S" component by sucrose gradient sedimentation. We identified two types of 5' terminals, m7G(5')pppNmpN and m7G(5')pppNm-pNmpN, in which the first methylated nucleoside is 7-methylguanosine, the second is either N6, 2'-O-dimethyladenosine, 2'-O-methyladenosine, or 2'-O-methylguanosine, and the third is 2'-O-methyluridine, 2'-O-methylcytidine, or 2'-O-methyladenosine. Approximately 1.7% of the 32P label was present in the 5' terminal structures. Assuming a similar specific radioactivity for all phosphates, this percentage corresponds to an average of one terminal per 335 nucleotides. Histone mRNA differed from bulk polyadenylylated mRNA of HeLa cells in lacking significant amounts of 2'-O-methyluridine or 2'-O-methylcytidine in the second position of the 5' terminal oligonucleotide and in lacking N6-methyladenosine residues at internal positions.