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Nucleic Acids Research, 1978, Vol. 5, No. 9 3237-3248
© 1978

Articles

Requirement for 7-methylguanosine in translation of globin mRNA in vivo

Raymond E. Lockard* and Charles Lane+

*Department of Biology, Massachusetts Institute of Technology Cambridge, MA 02139, USA +National Institute for Medical Research The Ridgeway, Mill Hill, London NW7 1AA, UK

Received May 16, 1978. The 7-methylguanosine (m7G residue present in tha m7G5' ppp5' X- "CAP" structure of rabbit globin mRNA was removed quantitatively by periodate oxidation followed by ß-elimination in the presence of cyclohexylamine. The RNA thus treated was intact and exhibited no signs of degradation as examined by polyacrylamide gel electrophoresis in formamide. Assay for protein synthesis using a wheat germ cell-free system showed that the globin mRNA lacking m7G had lost most of its messenger activity. Identical treatment, of satellite tobacco necrosis virus (STNV) RNA, which does not contain the 5 - terminal "CAP" structure, resulted in no loss of its mRNA activity. Since the importance of the m7G residue in eukaryotic mRNA has not yet been shown essential for translation in vivo, both untreated and treated globin mRNAs were injected into frog oocytes and their translation into globin was measured at intervals over a ninety-six hour period. Globin mRNA either treated with periodate alone or lacking in m7G altogether were both found to have lost more than 90% of their activity in vivo.


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