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Nucleic Acids Research, 1988, Vol. 16, No. 18 8953-8962
© 1988

Articles

ß-globin mRNAs capped with m7G, m22.7G or m32.2.7G differ in intrinsic translation efficiency

E. Darzynkiewicz1, J. Stepinski2, I. Ekiel3, Y. Jin+, D. Haber1, T. Sijuwade and S.M. Tahara*

Department of Microbiology, USC School of Medicine Los Angeles, CA 90033—1054, USA 1Department of Biophysics, Institute of Experimental Physics, University of Warsaw 02–089 Warsaw 2Department of Organic Chemistry, Institute of Chemistry, University of Warsaw 02-093 Warsaw, Poland 3Division of Biological Sciences, National Research Council of Canada Ottawa, Ontario, Canada

*To whom correspondence should be addressed

Received April 29, 1988. Revised August 22, 1988. Accepted August 22, 1988.

We examined the intramolecular effect of altered cap structures on translation efficiency of artificial ß-globin mRNAs. For these studies, synthetic dinucleotides of the form X(5')ppp(5')G [X=7-methyl guanosine (m7G), 2, 7-dimethyl guanosine (Formula) or 2,2,7-trimethyl guanosine (Formula), were transcriptionally Incorporated into mRNAs, containing rabbit ß-globin coding sequences, using T7 RNA potymerase and a ß-globin cDNA template. These synthetic mRNAs were assayed in reticulocyte lysate for activity relative to m7G-capped mRNA. Formula-Capped mRNA was found to be 1.5-fold more active than m7G-capped mRNA. Messenger RNA capped with Formula was less active with activity of 0.24 relative to its m7G-capped counterpart (activity = 1.0). These data suggest that m7G-capped mRNAs become more active as translation templates after addition of a single N2 methyl moiety, which is especially pertinent to gene expression in togaviridae. The latter are observed to synthesize Formula and Formula-capped mRNAs in addition to m7G-capped templates during the course of infection in animal cells.

+Permanent address: Shanghai Institute of Biochemistry, Academia Sinica, Shanghai, China


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