Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2

In yeast, Elongator-dependent tRNA modifications are regulated by the Kti11•Kti13 dimer and hijacked for cell killing by zymocin, a tRNase ribotoxin. Kti11 (alias Dph3) also controls modification of elongation factor 2 (EF2) with diphthamide, the target for lethal ADP-ribosylation by diphtheria toxi...

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Main Authors: Meike Arend, Koray Ütkür, Harmen Hawer, Klaus Mayer, Namit Ranjan, Lorenz Adrian, Ulrich Brinkmann, Raffael Schaffrath
Format: Article
Language:English
Published: Shared Science Publishers OG 2022-08-01
Series:Microbial Cell
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Online Access:http://microbialcell.com/researcharticles/2023a-arend-microbial-cell/
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author Meike Arend
Koray Ütkür
Harmen Hawer
Klaus Mayer
Namit Ranjan
Lorenz Adrian
Ulrich Brinkmann
Raffael Schaffrath
author_facet Meike Arend
Koray Ütkür
Harmen Hawer
Klaus Mayer
Namit Ranjan
Lorenz Adrian
Ulrich Brinkmann
Raffael Schaffrath
author_sort Meike Arend
collection DOAJ
description In yeast, Elongator-dependent tRNA modifications are regulated by the Kti11•Kti13 dimer and hijacked for cell killing by zymocin, a tRNase ribotoxin. Kti11 (alias Dph3) also controls modification of elongation factor 2 (EF2) with diphthamide, the target for lethal ADP-ribosylation by diphtheria toxin (DT). Diphthamide formation on EF2 involves four biosynthetic steps encoded by the DPH1-DPH7 network and an ill-defined KTI13 function. On further examining the latter gene in yeast, we found that kti13Δ null-mutants maintain unmodified EF2 able to escape ADP-ribosylation by DT and to survive EF2 inhibition by sordarin, a diphthamide-dependent antifungal. Consistently, mass spectrometry shows kti13Δ cells are blocked in proper formation of amino-carboxyl-propyl-EF2, the first diphthamide pathway intermediate. Thus, apart from their common function in tRNA modification, both Kti11/Dph3 and Kti13 share roles in the initiation step of EF2 modification. We suggest an alias KTI13/DPH8 nomenclature indicating dual-functionality analogous to KTI11/DPH3.
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issn 2311-2638
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publishDate 2022-08-01
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spelling doaj-art-cbab6fc1de5a40b2b53aba41197cf1862025-08-20T02:57:39ZengShared Science Publishers OGMicrobial Cell2311-26382022-08-0110919520310.15698/mic2023.09.804Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2Meike Arend0Koray Ütkür1Harmen Hawer2Klaus Mayer3Namit Ranjan4Lorenz Adrian5Ulrich Brinkmann6Raffael Schaffrath7Institute of Biology, Division of Microbiology, University of Kassel, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.Institute of Biology, Division of Microbiology, University of Kassel, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.Institute of Biology, Division of Microbiology, University of Kassel, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.Roche Pharma Research and Early Development, Large Molecule Research, Roche Innovation Center München, Nonnenwald 2, 82377 Penzberg, Germany.Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.Environmental Biotechnology, Helmholtz Centre for Environmental Research - UFZ, 04318 Leipzig, Germany.Roche Pharma Research and Early Development, Large Molecule Research, Roche Innovation Center München, Nonnenwald 2, 82377 Penzberg, Germany.Institute of Biology, Division of Microbiology, University of Kassel, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.In yeast, Elongator-dependent tRNA modifications are regulated by the Kti11•Kti13 dimer and hijacked for cell killing by zymocin, a tRNase ribotoxin. Kti11 (alias Dph3) also controls modification of elongation factor 2 (EF2) with diphthamide, the target for lethal ADP-ribosylation by diphtheria toxin (DT). Diphthamide formation on EF2 involves four biosynthetic steps encoded by the DPH1-DPH7 network and an ill-defined KTI13 function. On further examining the latter gene in yeast, we found that kti13Δ null-mutants maintain unmodified EF2 able to escape ADP-ribosylation by DT and to survive EF2 inhibition by sordarin, a diphthamide-dependent antifungal. Consistently, mass spectrometry shows kti13Δ cells are blocked in proper formation of amino-carboxyl-propyl-EF2, the first diphthamide pathway intermediate. Thus, apart from their common function in tRNA modification, both Kti11/Dph3 and Kti13 share roles in the initiation step of EF2 modification. We suggest an alias KTI13/DPH8 nomenclature indicating dual-functionality analogous to KTI11/DPH3.http://microbialcell.com/researcharticles/2023a-arend-microbial-cell/budding yeastef2 diphthamide modificationdiphtheria toxintrna modificationelongatortrnase zymocin
spellingShingle Meike Arend
Koray Ütkür
Harmen Hawer
Klaus Mayer
Namit Ranjan
Lorenz Adrian
Ulrich Brinkmann
Raffael Schaffrath
Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
Microbial Cell
budding yeast
ef2 diphthamide modification
diphtheria toxin
trna modification
elongator
trnase zymocin
title Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
title_full Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
title_fullStr Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
title_full_unstemmed Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
title_short Yeast gene KTI13 (alias DPH8) operates in the initiation step of diphthamide synthesis on elongation factor 2
title_sort yeast gene kti13 alias dph8 operates in the initiation step of diphthamide synthesis on elongation factor 2
topic budding yeast
ef2 diphthamide modification
diphtheria toxin
trna modification
elongator
trnase zymocin
url http://microbialcell.com/researcharticles/2023a-arend-microbial-cell/
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