Fungal Kti12 proteins display unusual linker regions and unique ATPase p-loops
Open Access
- 31 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Current Genetics
- Vol. 66 (4), 823-833
- https://doi.org/10.1007/s00294-020-01070-2
Abstract
Kti12 (Kluyveromyces lactis toxin insensitive 12) is an evolutionary highly conserved ATPase, crucial for the tRNA-modification activity of the eukaryotic Elongator complex. The protein consists of an N-terminal ATPase and a C-terminal tRNA-binding domain, which are connected by a flexible linker. The precise role of the linker region and its involvement in the communication between the two domains and their activities remain elusive. Here, we analyzed all available Kti12 protein sequences and report the discovery of a subset of Kti12 proteins with abnormally long linker regions. These Kti12 proteins are characterized by a co-occurring lysine to leucine substitution in their Walker A motif, previously thought to be invariable. We show that the K14L substitution lowers the affinity to ATP, but does not affect the catalytic activity of Kti12 at high ATP concentrations. We compare the activity of mutated variants of Kti12 in vitro with complementation assays in vivo in yeast. Ultimately, we compared Kti12 to other known p-loop ATPase family members known to carry a similar deviant Walker A motif. Our data establish Kti12 of Eurotiomycetes as an example of eukaryotic ATPase harboring a significantly deviating but still functional Walker A motif.Keywords
Funding Information
- Fundacja na rzecz Nauki Polskiej (TEAM TECH CORE FACILITY/2017-4/6, FirstTEAM/2016-1/2)
- Narodowe Centrum Nauki (UMO-2018/31/B/NZ1/03559)
- Deutsche Forschungsgemeinschaft (BR921/9)
This publication has 59 references indexed in Scilit:
- Elongator function in tRNA wobble uridine modification is conserved between yeast and plantsMolecular Microbiology, 2010
- Mammalian ALKBH8 Possesses tRNA Methyltransferase Activity Required for the Biogenesis of Multiple Wobble Uridine Modifications Implicated in Translational DecodingMolecular and Cellular Biology, 2010
- I-TASSER: a unified platform for automated protein structure and function predictionNature Protocols, 2010
- Protein structure prediction on the Web: a case study using the Phyre serverNature Protocols, 2009
- Mechanistic characterization of the sulfur-relay system for eukaryotic 2-thiouridine biogenesis at tRNA wobble positionsNucleic Acids Research, 2009
- Jalview Version 2—a multiple sequence alignment editor and analysis workbenchBioinformatics, 2009
- Characterization and evolutionary history of an archaeal kinase involved in selenocysteinyl-tRNA formationNucleic Acids Research, 2008
- Interactive Tree Of Life (iTOL): an online tool for phylogenetic tree display and annotationBioinformatics, 2006
- Mutations in ABO1/ELO2, a Subunit of Holo-Elongator, Increase Abscisic Acid Sensitivity and Drought Tolerance in Arabidopsis thalianaMolecular and Cellular Biology, 2006
- MUSCLE: multiple sequence alignment with high accuracy and high throughputNucleic Acids Research, 2004