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Title: Structural and functional characterization of KEOPS dimerization by Pcc1 and its role in t6A biosynthesis

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkw542· OSTI ID:1328017
 [1];  [2];  [1];  [3];  [3];  [2];  [4];  [1];  [1]
  1. Lunenfeld-Tanenbaum Research Inst., Toronto, ON (Canada); Univ. of Toronto, ON (Canada)
  2. McMaster Univ., Hamilton, ON (Canada)
  3. Univ. of Toronto, ON (Canada)
  4. Cornell Univ., Argonne, IL (United States)
+ Show Author Affiliations

KEOPS is an ancient protein complex required for the biosynthesis of N6-threonylcarbamoyladenosine (t6A), a universally conserved tRNA modification found on all ANN-codon recognizing tRNAs. KEOPS consist minimally of four essential subunits, namely the proteins Kae1, Bud32, Cgi121 and Pcc1, with yeast possessing the fifth essential subunit Gon7. Bud32, Cgi121, Pcc1 and Gon7 appear to have evolved to regulate the central t6A biosynthesis function of Kae1, but their precise function and mechanism of action remains unclear. Pcc1, in particular, binds directly to Kae1 and by virtue of its ability to form dimers in solution and in crystals, Pcc1 was inferred to function as a dimerization module for Kae1 and therefore KEOPS. We now present a 3.4 Å crystal structure of a dimeric Kae1–Pcc1 complex providing direct evidence that Pcc1 can bind and dimerize Kae1. Further biophysical analysis of a complete archaeal KEOPS complex reveals that Pcc1 facilitates KEOPS dimerization in vitro. Interestingly, while Pcc1-mediated dimerization of KEOPS is required to support the growth of yeast, it is dispensable for t6A biosynthesis by archaeal KEOPS in vitro, raising the question of how precisely Pcc1-mediated dimerization impacts cellular biology.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
Canadian Inst. of Health Research (CIHR) Foundation; National Inst. of Health; NIH-ORIP HEI
Grant/Contract Number:
AC02-06CH11357; FDN 143277; FDN 143343; P41 GM103403; S10 RR029205
OSTI ID:
1328017
Journal Information:
Nucleic Acids Research, Vol. 44, Issue 14; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 16 works
Citation information provided by
Web of Science

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Cited By (8)

Defects in t6A tRNA modification due to GON7 and YRDC mutations lead to Galloway-Mowat syndrome journal September 2019
Transfer RNA Modification Enzymes from Thermophiles and Their Modified Nucleosides in tRNA journal October 2018
Molecular basis for t6A modification in human mitochondria journal February 2020
Structure of a reaction intermediate mimic in t6A biosynthesis bound in the active site of the TsaBD heterodimer from Escherichia coli journal February 2021
Proteomic analysis of the human KEOPS complex identifies C14ORF142 as a core subunit homologous to yeast Gon7 journal November 2016
The structure of the TsaB/TsaD/TsaE complex reveals an unexpected mechanism for the bacterial t6A tRNA-modification journal August 2019
Structure and Mechanism of a Bacterial t6A Biosynthesis System journal April 2018
Identifying direct contacts between protein complex subunits from their conditional dependence in proteomics datasets journal October 2017

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