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Nucleic Acids Research, 1994, Vol. 22, No. 3 419-426
© 1994

RNA

Context effects on N6-adenosine methylation sites in prolactin mRNA

Prema Narayan+, Rachael L. Ludwiczak, Edward C. Goodwin and Fritz M. Rottman*

Department of Molecular Biology and Microbiology, Case Western Reserve University Cleveland, OH 44106, USA

*To whom correspondence should be addressed

Received September 17, 1993. Accepted December 28, 1993.

The methylatlon of internal adenosine residues in mRNA only occurs within GAC or AAC sequences. Although both of these sequence motifs are utilized, a general preference has been noted for the extended sequence RGACU. Not all RGACU sequences in an mRNA are methylated and the mechanisms that govern the selection of methylatlon sites In mRNA remain unclear. To address this problem we have examined the methylatlon of transcripts containing sequences of a natural mRNA, namely, bovine prolactln mRNA. In this mRNA, a specific AGACU sequence in the 3' untranslated region Is the predominant site of methylatlon both in vivo and in vitro. The degree to which N6-adenoslne methyltransferase recognizes the sequence context of the consensus methylatlon site was explored by mutatlonal analysis of the nucleotIdes adjacent to the core sequence as well as the extended regions in which the core element was found. Our results Indicate that efficient methylatlon depends on the extended five nucleotide consensus sequence but is strongly influenced by the context in which the consensus sequence occurs within the overall mRNA molecule. Furthermore, consensus methylatlon sites present in an RNA duplex are not recognized by the methyltransferase.

+Present address: Department of Biochemistry, Life Sciences Building, University of Georgia, Athens, GA 30602, USA


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