Positioning of the mRNA stop signal with respect to polypeptide chain release factors and ribosomal proteins in 80S ribosomes

FEBS Letters
Konstantin N BulyginLev L Kisselev

Abstract

To study positioning of the mRNA stop signal with respect to polypeptide chain release factors (RFs) and ribosomal components within human 80S ribosomes, photoreactive mRNA analogs were applied. Derivatives of the UUCUAAA heptaribonucleotide containing the UUC codon for Phe and the stop signal UAAA, which bore a perfluoroaryl azido group at either the fourth nucleotide or the 3'-terminal phosphate, were synthesized. The UUC codon was directed to the ribosomal P site by the cognate tRNA(Phe), targeting the UAA stop codon to the A site. Mild UV irradiation of the ternary complexes consisting of the 80S ribosome, the mRNA analog and tRNA resulted in tRNA-dependent crosslinking of the mRNA analogs to the 40S ribosomal proteins and the 18S rRNA. mRNA analogs with the photoreactive group at the fourth uridine (the first base of the stop codon) crosslinked mainly to protein S15 (and much less to S2). For the 3'-modified mRNA analog, the major crosslinking target was protein S2, while protein S15 was much less crosslinked. Crosslinking of eukaryotic (e) RF1 was entirely dependent on the presence of a stop signal in the mRNA analog. eRF3 in the presence of eRF1 did not crosslink, but decreased the yield of eRF1 crosslinking. We conclude...Continue Reading

References

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Nov 21, 2000·Trends in Biochemical Sciences·L L Kisselev, R H Buckingham

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Citations

Aug 30, 2011·The Journal of Biological Chemistry·Zofia M A Chrzanowska-LightowlersRobert N Lightowlers
Jan 7, 2003·The EMBO Journal·Lev KisselevLudmila Frolova
Nov 12, 2005·Nucleic Acids Research·Petr KolosovLev Kisselev
Mar 27, 2007·Molekuliarnaia biologiia·Iu N Vorob'ev, L L Kiselev
Jul 10, 2008·Molekuliarnaia biologiia·Iu S KhaĭrulinaG G Karpova
Dec 16, 2003·Proceedings of the National Academy of Sciences of the United States of America·Eric JanPeter Sarnow
Jun 2, 2005·Reviews of Physiology, Biochemistry and Pharmacology·S RospertY Dubaquié
Sep 7, 2007·Molecular Cell·Hamid Reza Soleimanpour-LichaeiZofia Chrzanowska-Lightowlers
Jan 20, 2012·Acta Biochimica Et Biophysica Sinica·Dmitri Graifer, Galina Karpova
Nov 30, 2013·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Dmitri Graifer, Galina Karpova
Sep 25, 2007·Bioorganicheskaia khimiia·M V MolotkovG G Karpova
Jan 4, 2020·Journal of Cell Science·Shreya Ahana AyyubZofia M Chrzanowska-Lightowlers
Nov 6, 2004·The Biochemical Journal·Dmitri GraiferGalina Karpova

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