Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator

Mehlgarten, Constance; Prochaska, Heike; Hammermeister, Alexander; Abdel-Fattah, Wael; Wagner, Melanie; Krutyhołowa, Rościsław; Jun, Sang Eun; Kim, Gyung-Tae; Glatt, Sebastian; Breunig, Karin D.; Stark, Michael J. R.; Schaffrath, Raffael

doi:10.3390/toxins9090272

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Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator

Constance Mehlgarten

¹,

Heike Prochaska

¹,

Alexander Hammermeister

Wael Abdel-Fattah

Melanie Wagner

¹,

Rościsław Krutyhołowa

^3,4

⁵,

⁵,

¹,

⁶

and

Raffael Schaffrath

^1,2,*

Institut für Biologie, Martin Luther Universität Halle-Wittenberg, Weinbergweg 10, 06120 Halle/Saale, Germany

Institut für Biologie, FG Mikrobiologie, Universität Kassel, Heirich-Plett-Str. 40, 34132 Kassel, Germany

Max Planck Research Group at the Malopolska Centre of Biotechnology, Jagiellonian University, 31-007 Krakow, Poland

⁴

Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 31-007 Krakow, Poland

⁵

Department of Molecular Biotechnology, Dong-A University, Busan 604-714, Korea

⁶

Centre for Gene Regulation & Expression, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK

Author to whom correspondence should be addressed.

Academic Editor: Manfred J. Schmitt

Toxins 2017, 9(9), 272; https://doi.org/10.3390/toxins9090272

Received: 31 July 2017 / Revised: 29 August 2017 / Accepted: 3 September 2017 / Published: 5 September 2017

(This article belongs to the Special Issue Yeast Killer Toxins)

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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. View Full-Text

Keywords: zymocin; ribotoxin; tRNase; Kti12; Elongator complex; tRNA anticodon modification

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MDPI and ACS Style

Mehlgarten, C.; Prochaska, H.; Hammermeister, A.; Abdel-Fattah, W.; Wagner, M.; Krutyhołowa, R.; Jun, S.E.; Kim, G.-T.; Glatt, S.; Breunig, K.D.; Stark, M.J.R.; Schaffrath, R. Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator. Toxins 2017, 9, 272. https://doi.org/10.3390/toxins9090272

AMA Style

Mehlgarten C, Prochaska H, Hammermeister A, Abdel-Fattah W, Wagner M, Krutyhołowa R, Jun SE, Kim G-T, Glatt S, Breunig KD, Stark MJR, Schaffrath R. Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator. Toxins. 2017; 9(9):272. https://doi.org/10.3390/toxins9090272

Chicago/Turabian Style

Mehlgarten, Constance, Heike Prochaska, Alexander Hammermeister, Wael Abdel-Fattah, Melanie Wagner, Rościsław Krutyhołowa, Sang E. Jun, Gyung-Tae Kim, Sebastian Glatt, Karin D. Breunig, Michael J.R. Stark, and Raffael Schaffrath. 2017. "Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator" Toxins 9, no. 9: 272. https://doi.org/10.3390/toxins9090272

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Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator

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