Polyamine stimulation of eEF1A synthesis based on the unusual position of a complementary sequence to 18S rRNA in eEF1A mRNA
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It is thought that Shine–Dalgarno-like sequences, which exhibit complementarity to the nucleotide sequences at the 3′-end of 18S rRNA, are not present in eukaryotic mRNAs. However, complementary sequences consisting of more than 5 nucleotides to the 3′-end of 18S rRNA, i.e., a CR sequence, are present at −17 to −32 upstream from the initiation codon AUG in 18 mRNAs involved in protein synthesis except eEF1A mRNA. Thus, effects of the CR sequence in mRNAs and polyamines on protein synthesis were examined using control and polyamine-reduced FM3A and NIH3T3 cells. Polyamines did not stimulate protein synthesis encoded by 18 mRNAs possessing a normal CR sequence. When the CR sequence was deleted, protein synthetic activities decreased to less than 70 % of intact mRNAs. In eEF1A mRNA, the CR sequence was located at −33 to −39 upstream from the initiation codon AUG, and polyamines stimulated eEF1A synthesis about threefold. When the CR sequence was shifted to −22 to −28 upstream from the AUG, eEF1A synthesis increased in polyamine-reduced cells and the degree of polyamine stimulation decreased greatly. The results indicate that the CR sequence exists in many eukaryotic mRNAs, and the location of a CR sequence in mRNAs influences polyamine stimulation of protein synthesis.
KeywordsPolyamines Protein synthesis mRNA 5′-UTR CR sequence
- CR sequence
Complementary sequence to 18S rRNA
5′-Untranslated region of mRNA
We thank Drs. A. J. Michael and K. Williams for their help in preparing the manuscript. We also thank Dr. R. G. Mirmira for his kind gift of antibody against hypusinated eIF5A. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
Conflict of interest
The authors declare that they have no conflict of interest.
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