Preprint Communication Version 1 Preserved in Portico This version is not peer-reviewed

Use of a Yeast TRNase Killer Toxin to Diagnose Kti12 Motifs Required for TRNA Modification by Elongator

Raffael Schaffrath
* ORCID logo ,
Constance Mehlgarten
,
Heike Prochaska
,
Alexander Hammermeister
,
Wael Abdel-Fattah
ORCID logo ,
Melanie Wagner
,
Rościsław Krutyhołowa
,
Sang Eun Jun
,
Gyung-Tae Kim
,
Sebastian Glatt
ORCID logo ,
Karin D. Breunig
,
Michael J.R. Stark
ORCID logo
Version 1 : Received: 31 July 2017 / Approved: 2 August 2017 / Online: 2 August 2017 (11:45:15 CEST)

How to cite: Schaffrath, R.; Mehlgarten, C.; Prochaska, H.; Hammermeister, A.; Abdel-Fattah, W.; Wagner, M.; Krutyhołowa, R.; Jun, S.E.; Kim, G.; Glatt, S.; Breunig, K.D.; Stark, M.J. Use of a Yeast TRNase Killer Toxin to Diagnose Kti12 Motifs Required for TRNA Modification by Elongator. Preprints 2017, 2017080001. https://doi.org/10.20944/preprints201708.0001.v1 Schaffrath, R.; Mehlgarten, C.; Prochaska, H.; Hammermeister, A.; Abdel-Fattah, W.; Wagner, M.; Krutyhołowa, R.; Jun, S.E.; Kim, G.; Glatt, S.; Breunig, K.D.; Stark, M.J. Use of a Yeast TRNase Killer Toxin to Diagnose Kti12 Motifs Required for TRNA Modification by Elongator. Preprints 2017, 2017080001. https://doi.org/10.20944/preprints201708.0001.v1

Abstract

Saccharomyces cerevisiae cells are killed by zymocin, a tRNase ribotoxin complex from Kluyveromyces lactis, which cleaves anticodons and inhibits protein synthesis. Zymocin’s action requires specific chemical modification of uridine bases in the anticodon wobble position (U34) by the Elongator complex (Elp1-Elp6). Hence, loss of anticodon modification in mutants lacking Elongator or related KTI (K. lactis Toxin Insensitive) genes protects against tRNA cleavage and confers resistance to the toxin. Here, we show that zymocin can be used as a tool to genetically analyse KTI12, a gene previously shown to code for an Elongator partner protein. From a kti12 mutant pool of zymocin survivors, we identify motifs in Kti12 that are functionally directly coupled to Elongator activity. In addition, shared requirement of U34 modifications for nonsense and missense tRNA suppression (SUP4; SOE1) strongly suggests that Kti12 and Elongator cooperate to assure proper tRNA functioning. We show that the Kti12 motifs are conserved in plant ortholog DRL1/ELO4 from Arabidopsis thaliana and seem to be involved in binding of cofactors (e.g. nucleotides, calmodulin). Elongator interaction defects triggered by mutations in these motifs correlate with phenotypes typical for loss of U34 modification. Thus, tRNA modification by Elongator appears to require physical contact with Kti12, and our preliminary data suggest that metabolic signals may affect proper communication between them.

Keywords

Zymocin; ribotoxin; tRNase; Kti12; Elongator complex; tRNA anticodon modification

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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