en
Scientific article
Open access
English

Structural and functional studies of Arabidopsis thaliana triphosphate tunnel metalloenzymes reveal roles for additional domains

Published inJournal of biological chemistry, vol. 298, no. 11, 102438
Publication date2022-11
First online date2022-08-30
Abstract

Triphosphate tunnel metalloenzymes (TTMs) are found in all biological kingdoms and have been characterized in microorganisms and animals. Members of the TTM family have divergent biological functions and act on a range of triphosphorylated substrates (RNA, thiamine triphosphate, and inorganic polyphosphate). TTMs in plants have received considerably less attention and are unique in that some homologs harbor additional domains including a P-loop kinase and transmembrane domain. Here, we report on structural and functional aspects of the multimodular TTM1 and TTM2 of Arabidopsis thaliana. Our tissue and cellular microscopy studies show that both AtTTM1 and AtTTM2 are expressed in actively dividing (meristem) tissue and are tail-anchored proteins at the outer mitochondrial membrane, mediated by the single C-terminal transmembrane domain, supporting earlier studies. In addition, we reveal from crystal structures of AtTTM1 in the presence and absence of a nonhydrolyzable ATP analog a catalytically incompetent TTM tunnel domain tightly interacting with the P-loop kinase domain that is locked in an inactive conformation. Our structural comparison indicates that a helical hairpin may facilitate movement of the TTM domain, thereby activating the kinase. Furthermore, we conducted genetic studies to show that AtTTM2 is important for the developmental transition from the vegetative to the reproductive phase in Arabidopsis, whereas its closest paralog AtTTM1 is not. We demonstrate through rational design of mutations based on the 3D structure that both the P-loop kinase and TTM tunnel modules of AtTTM2 are required for the developmental switch. Together, our results provide insight into the structure and function of plant TTM domains.

eng
Keywords
  • Arabidopsis
  • X-ray crystallography
  • Development
  • Mitochondria
  • Triphosphate tunnel metalloenzyme
  • Uridine/cytidine kinase
  • Animals
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / metabolism
  • Polyphosphates
  • Metalloproteins / chemistry
  • Acid Anhydride Hydrolases / metabolism
Citation (ISO format)
PESQUERA-ALONSO, Marta et al. Structural and functional studies of Arabidopsis thaliana triphosphate tunnel metalloenzymes reveal roles for additional domains. In: Journal of biological chemistry, 2022, vol. 298, n° 11, p. 102438. doi: 10.1016/j.jbc.2022.102438
Main files (1)
Article (Published version)
Secondary files (1)
Identifiers
ISSN of the journal0021-9258
19views
34downloads

Technical informations

Creation05/15/2023 10:30:13 AM
First validation05/15/2023 10:52:59 AM
Update time05/15/2023 10:52:59 AM
Status update05/15/2023 10:52:59 AM
Last indexation05/06/2024 3:52:06 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack