Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN.
AtHESPERIN
circadian rhythms
deadenylation
mRNA decay
poly(A) tail
Journal
FEBS open bio
ISSN: 2211-5463
Titre abrégé: FEBS Open Bio
Pays: England
ID NLM: 101580716
Informations de publication
Date de publication:
05 2022
05 2022
Historique:
revised:
04
10
2020
received:
01
05
2020
accepted:
20
10
2020
pubmed:
24
10
2020
medline:
6
5
2022
entrez:
23
10
2020
Statut:
ppublish
Résumé
The 24-h molecular clock is based on the stability of rhythmically expressed transcripts. The shortening of the poly(A) tail of mRNAs is often the first and rate-limiting step that determines the lifespan of a mRNA and is catalyzed by deadenylases. Herein, we determine the catalytic site of Hesperin, a recently described circadian deadenylase in plants, using a modified site-directed mutagenesis protocol and a custom vector, pATHRA. To explore the catalytic efficiency of AtHESPERIN, we investigated the effect of AMP and neomycin, and used molecular modeling simulations to propose a catalytic mechanism. Collectively, the biochemical and in silico results classify AtHESPERIN in the exonuclease-endonuclease-phosphatase deadenylase superfamily and contribute to the understanding of the intricate mechanisms of circadian mRNA turnover.
Identifiants
pubmed: 33095977
doi: 10.1002/2211-5463.13011
pmc: PMC9063446
doi:
Substances chimiques
RNA, Messenger
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1036-1049Subventions
Organisme : University of Thessaly Research Committee
ID : 3439
Organisme : University of Thessaly Research Committee
ID : 3817
Organisme : University of Thessaly Research Committee
ID : 5851
Organisme : OMIC-ENGINE (MIS 5002636) implemented under "Reinforcement of the Research and Innovation Infrastructure", Operational Programme "Competitiveness, Entrepreneurship and Innovation" (NSRF 2014-2020)
ID : MIS 5002636
Organisme : State Scholarships Foundation (IKY) Fellowship of Excellence for Postgraduate Studies in Greece - Siemens Programme (to AK)
Organisme : European Regional Development Fund
Organisme : National Research Foundation
Informations de copyright
© 2020 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.
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