Thiamine metabolism genes in diatoms are not regulated by thiamine despite the presence of predicted riboswitches.
Phaeodactylum tricornutum
CRISPR/Cas9
TPP riboswitch
aptamer prediction
diatoms
thiamine biosynthesis
thiamine uptake
Journal
The New phytologist
ISSN: 1469-8137
Titre abrégé: New Phytol
Pays: England
ID NLM: 9882884
Informations de publication
Date de publication:
09 2022
09 2022
Historique:
received:
03
01
2022
accepted:
20
05
2022
pubmed:
3
6
2022
medline:
4
8
2022
entrez:
2
6
2022
Statut:
ppublish
Résumé
Thiamine pyrophosphate (TPP), an essential co-factor for all species, is biosynthesised through a metabolically expensive pathway regulated by TPP riboswitches in bacteria, fungi, plants and green algae. Diatoms are microalgae responsible for c. 20% of global primary production. They have been predicted to contain TPP aptamers in the 3'UTR of some thiamine metabolism-related genes, but little information is known about their function and regulation. We used bioinformatics, antimetabolite growth assays, RT-qPCR, targeted mutagenesis and reporter constructs to test whether the predicted TPP riboswitches respond to thiamine supplementation in diatoms. Gene editing was used to investigate the functions of the genes with associated TPP riboswitches in Phaeodactylum tricornutum. We found that thiamine-related genes with putative TPP aptamers are not responsive to supplementation with thiamine or its precursor 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP), and targeted mutation of the TPP aptamer in the THIC gene encoding HMP-P synthase does not deregulate thiamine biosynthesis in P. tricornutum. Through genome editing we established that PtTHIC is essential for thiamine biosynthesis and another gene, PtSSSP, is necessary for thiamine uptake. Our results highlight the importance of experimentally testing bioinformatic aptamer predictions and provide new insights into the thiamine metabolism shaping the structure of marine microbial communities with global biogeochemical importance.
Identifiants
pubmed: 35653609
doi: 10.1111/nph.18296
pmc: PMC9544697
doi:
Substances chimiques
Riboswitch
0
Thiamine Pyrophosphate
Q57971654Y
Thiamine
X66NSO3N35
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1853-1867Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/L002957/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/M011194/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/M018180/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/R021694/1
Pays : United Kingdom
Informations de copyright
© 2022 The Authors. New Phytologist © 2022 New Phytologist Foundation.
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