Complex of transfer-messenger RNA and elongation factor Tu: Unexpected modes of interaction

Maria I. Zvereva, Pavel V. Ivanov, Yoshika Teraoka, Natalya I. Topilina, Olga A. Dontsova, Alexey A. Bogdanov, Markus Kalkum, Knud H. Nierhaus, Olga V. Shpanchenko

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)

Abstract

Transfer-messenger RNA (tmRNA) is a stable RNA in bacteria of 360 ± 40 nucleotides that can be charged with alanine and can function as both tRNA and mRNA. Ribosomes that are stalled either in a coding region of mRNA or at the 3′ end of an mRNA fragment lacking a stop codon are rescued by replacing their mRNA for tmRNA. Here we demonstrate that the interaction of tmRNA with the elongation factor Tu shows unexpected features. Deacylated tmRNA can form a complex with either EF-Tu·GDP or EF-Tu·GTP, the association constants are about one order of magnitude smaller than that of an Ala-tRNA·EF-Tu·GTP complex. tmRNA as well as Ala-tmRNA can be efficiently cross-linked with EF-Tu·GDP using a zero-length cross-link. The efficiency of cross-linking in the case of deacylated tmRNA does not depend on an intact CCA-3′ end and is about the same, regardless whether protein mixtures such as the post-ribosomal supernatant (S100 enzymes) or purified EF-Tu are present. Two cross-linking sites with EF-Tu·GDP have been identified that are located outside the tRNA part of tmRNA, indicating an unusual interaction of tmRNA with EF-Tu·GDP.

Original languageEnglish
Pages (from-to)47702-47708
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number50
DOIs
Publication statusPublished - 14 Dec 2001
Externally publishedYes

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