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Nucleic Acids Research, 1984, Vol. 12, No. 23 9205-9208
© 1984

MOLECULAR BIOLOGY

Partial methylation of two adjacent adenosines in ribosomes from Euglena gracilis chloroplasts suggests evolutionary loss of an intermediate stage in the methyl-transfer reaction

C.P.J.J. Van Buul, M. Hamersma, W. Visser and P.H. Van Knippenberg

Department of Biochemistry, State University of Leiden P.O.Box 9503, 2300 RA Leiden, The Netherlands

Received July 19, 1984. Revised November 7, 1984. Accepted November 7, 1984.

Bacterial, cytoplasmic and organellar ribosomes from a wide phylogenetic spectrum of organisms have a characteristic m26Am26A structure near the 3' end of the RNA of the small ribosomal subunit (SSU). We have studied one of the few exceptions to this extremely conserved post-transcriptionally modified sequence, i.e. dimethylation of only one of the two A's in chloroplasts from Euglena gracilis. It was established that only the A closest to the 5' end is dimethylated, the other one being unmodified.

The methylation reaction was studied in vitro using ribosomes from a kasugamycin resistant mutant (KsgA) of Escherichia coli and purified methyl-transferase. Using limited amounts of the methyldonor S-adenosylmethionine (SAM) a partial level of methylation (50% of control) was attained. It is shown that in this case the 3' proximal A is dimethylated while the other is not. This suggests that dimethylation takes place in two successive stages. Apparently in E. gracilis chloroplasts the first stage of methylation does not occur.


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