Kluyveromyces lactis γ-toxin, a ribonuclease that recognizes the anticodon stem loop of tRNA

Jian Lu, Anders Esberg, Bo Huang, Anders S. Byström

Research output: Contribution to journalArticle

Abstract

Kluyveromyces lactis γ-toxin is a tRNA endonuclease that cleaves Saccharomyces cerevisiae tRNA mcm5s2UUC Glu3, tRNA mcm5s2UUU Lys and tRNA mcm5s2UUG Gln between position 34 and position 35. All three substrate tRNAs carry a 5-methoxycarbonylmethyl-2-thiouridine (mcm 5 s 2U) residue at position 34 (wobble position) of which the mcm 5 group is required for efficient cleavage. However, the different cleavage efficiencies of mcm 5s 2 U 34-containing tRNAs suggest that additional features of these tRNAs affect cleavage. In the present study, we show that a stable anticodon stem and the anticodon loop are the minimal requirements for cleavage by γ-toxin. A synthetic minihelix RNA corresponding to the anticodon stem loop (ASL) of the natural substrate tRNA mcm5s2UUC Glu3 is cleaved at the same position as the natural substrate. In ASL UUC Glu3, the nucleotides U 34U 35C 36A 37 C 38 are required for optimal γ-toxin cleavage, whereas a purine at position 32 or a G in position 33 dramatically reduces the cleavage of the ASL. Comparing modified and partially modified forms of E. coli and yeast tRNA UUC Glu reinforced the strong stimulatory effects of the mcm 5 group, revealed a weak positive effect of the s 2 group and a negative effect of the bacterial 5-methylaminomethyl (mnm 5) group. The data underscore the high specificity of this yeast tRNA toxin.

Original languageEnglish (US)
Pages (from-to)1072-1080
Number of pages9
JournalNucleic Acids Research
Volume36
Issue number4
DOIs
StatePublished - Mar 2008
Externally publishedYes

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Anticodon
Ribonucleases
Transfer RNA
RNA, Transfer, Gln
RNA, Transfer, Glu
Yeasts
RNA, Transfer, Lys
Endonucleases
zymocin
Saccharomyces cerevisiae
Nucleotides
RNA
Escherichia coli

ASJC Scopus subject areas

  • Genetics

Cite this

Kluyveromyces lactis γ-toxin, a ribonuclease that recognizes the anticodon stem loop of tRNA. / Lu, Jian; Esberg, Anders; Huang, Bo; Byström, Anders S.

In: Nucleic Acids Research, Vol. 36, No. 4, 03.2008, p. 1072-1080.

Research output: Contribution to journalArticle

Lu, Jian ; Esberg, Anders ; Huang, Bo ; Byström, Anders S. / Kluyveromyces lactis γ-toxin, a ribonuclease that recognizes the anticodon stem loop of tRNA. In: Nucleic Acids Research. 2008 ; Vol. 36, No. 4. pp. 1072-1080.
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