Structural changes of yeast tRNA(Tyr) caused by the binding of divalent ions in the presence of spermine.
The existence of specific sites in tRNA for the binding of divalent cations has been seriously questioned by electrostatic considerations [Leroy & Guéron (1979) Biopolymers, 16, 2429-2446]. However, our earlier studies of the binding of Mg2+ and Mn2+ to yeast tRNA(Tyr) have indicated that spermine creates new binding sites for divalent cations [Weygand-Durasević et al. (1977) Biochim. Biophys, Acta, 479, 332-344; Nöthig-Laslo et al. (1981) Eur. J. Biochem. 117, 263-267]. We have now used yeast tRNA(Tyr), spin labeled at the hypermodified purine (i6A-37) in the anticodon loop, to study the effect of spermine on the binding of manganese ions. The presence of eight spermine molecules per tRNA(Tyr) at high ionic strength (0.2 M NaCl, 0.05 M triethanolamine.HCl) and at low temperature (7 degrees C) enhances the binding of manganese to tRNA(Tyr). This effect could not be explained by electrostatic binding. The initial binding of manganese to tRNA(Tyr) affects the motional properties of the spin label indicating a change of the conformation of the anticodon loop. From the absence of the paramagnetic effect of manganese on the ESR spectra of the spin label one can conclude that the first binding site for manganese is at a distance from i6A-37, influencing the spin label motion through a long-range effect. The enhancement of the binding of manganese to tRNA(Tyr) by spermine is lost upon destruction of its specific macromolecular structure and it does not occur in single stranded or in double-stranded polynucleotides. The observed effect can be explained by the binding of Mn2+ to new sites, created by the binding of spermine, which are specific for the macromolecular structure of tRNA.
Nöthig-Laslo
V
V
Rudjer Bosković Institute, Zagreb, Yugoslavia.
Weygand-Durasević
I
I
Kućan
Z
Z
D000926:Anticodon; D002413:Cations, Divalent; D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D008345:Manganese / Q000378:metabolism*; D009690:Nucleic Acid Conformation / Q000187:drug effects; D012331:RNA, Fungal / Q000378:metabolism*; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012365:RNA, Transfer, Tyr / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013096:Spermine / Q000494:pharmacology*
D000926:Anticodon; D002413:Cations, Divalent; D012331:RNA, Fungal; D012345:RNA, Transfer, Amino Acid-Specific; D012365:RNA, Transfer, Tyr; D013096:Spermine; D008345:Manganese