The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis

Folates are indispensable for plant development, but their molecular mode of action remains elusive. We synthesized a probe, “5-F-THF-Dayne,” comprising 5-formyl-tetrahydrofolate (THF) coupled to a photoaffinity tag. Exploiting this probe in an affinity proteomics study in Arabidopsis thaliana, we r...

Full description

Bibliographic Details
Authors: Li, Weichao, Liang, Qiuju, Mishra, Ratnesh, Sánchez Muñoz, Raúl|||0000-0003-0235-2985, Wang, Huan, Chen, Xin, Van Der Straeten, Dominique, Zhang, Chunyi, Xiao, Youli
Format: article
Publication Date:2021
Country:España
Institution:Universitat Politècnica de Catalunya (UPC)
Repository:UPCommons. Portal del coneixement obert de la UPC
Language:English
OAI Identifier:oai:upcommons.upc.edu:2117/446414
Online Access:https://hdl.handle.net/2117/446414
https://dx.doi.org/10.1093/plcell/koab198
Access Level:Open access
Keyword:Folates
Folate biosynthesis
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetal
id ES_537c240b7dcfca1f99ef17ca45f0f963
oai_identifier_str oai:upcommons.upc.edu:2117/446414
network_acronym_str ES
network_name_str España
repository_id_str
spelling The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesisLi, WeichaoLiang, QiujuMishra, RatneshSánchez Muñoz, Raúl|||0000-0003-0235-2985Wang, HuanChen, XinVan Der Straeten, DominiqueZhang, ChunyiXiao, YouliFolatesFolate biosynthesisÀrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetalFolates are indispensable for plant development, but their molecular mode of action remains elusive. We synthesized a probe, “5-F-THF-Dayne,” comprising 5-formyl-tetrahydrofolate (THF) coupled to a photoaffinity tag. Exploiting this probe in an affinity proteomics study in Arabidopsis thaliana, we retrieved 51 hits. Thirty interactions were independently validated with in vitro expressed proteins to bind 5-F-THF with high or low affinity. Interestingly, the interactors reveal associations beyond one-carbon metabolism, covering also connections to nitrogen (N) metabolism, carbohydrate metabolism/photosynthesis, and proteostasis. Two of the interactions, one with the folate biosynthetic enzyme DIHYDROFOLATE REDUCTASE-THYMIDYLATE SYNTHASE 1 (AtDHFR-TS1) and another with N metabolism-associated glutamine synthetase 1;4 (AtGLN1;4), were further characterized. In silico and experimental analyses revealed G35/K36 and E330 as key residues for the binding of 5-F-THF in AtDHFR-TS1 and AtGLN1;4, respectively. Site-directed mutagenesis of AtGLN1;4 E330, which co-localizes with the ATP-binding pocket, abolished 5-F-THF binding as well as AtGLN1;4 activity. Furthermore, 5-F-THF was noted to competitively inhibit the activities of AtDHFR-TS1 and AtGLN1;4. In summary, we demonstrated a regulatory role for 5-F-THF in N metabolism, revealed 5-F-THF-mediated feedback regulation of folate biosynthesis, and identified a total of 14 previously unknown high-affinity binding cellular targets of 5-F-THF. Together, this sets a landmark toward understanding the role of folates in plant development.20212021-10-1120252025-11-17journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/446414https://dx.doi.org/10.1093/plcell/koab198reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/4464142026-05-27T15:37:01Z
dc.title.none.fl_str_mv The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
title The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
spellingShingle The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
Li, Weichao
Folates
Folate biosynthesis
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetal
title_short The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
title_full The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
title_fullStr The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
title_full_unstemmed The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
title_sort The 5-formyl-tetrahydrofolate proteome links folates with C/N metabolism and reveals feedback regulation of folate biosynthesis
dc.creator.none.fl_str_mv Li, Weichao
Liang, Qiuju
Mishra, Ratnesh
Sánchez Muñoz, Raúl|||0000-0003-0235-2985
Wang, Huan
Chen, Xin
Van Der Straeten, Dominique
Zhang, Chunyi
Xiao, Youli
author Li, Weichao
author_facet Li, Weichao
Liang, Qiuju
Mishra, Ratnesh
Sánchez Muñoz, Raúl|||0000-0003-0235-2985
Wang, Huan
Chen, Xin
Van Der Straeten, Dominique
Zhang, Chunyi
Xiao, Youli
author_role author
author2 Liang, Qiuju
Mishra, Ratnesh
Sánchez Muñoz, Raúl|||0000-0003-0235-2985
Wang, Huan
Chen, Xin
Van Der Straeten, Dominique
Zhang, Chunyi
Xiao, Youli
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Folates
Folate biosynthesis
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetal
topic Folates
Folate biosynthesis
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetal
description Folates are indispensable for plant development, but their molecular mode of action remains elusive. We synthesized a probe, “5-F-THF-Dayne,” comprising 5-formyl-tetrahydrofolate (THF) coupled to a photoaffinity tag. Exploiting this probe in an affinity proteomics study in Arabidopsis thaliana, we retrieved 51 hits. Thirty interactions were independently validated with in vitro expressed proteins to bind 5-F-THF with high or low affinity. Interestingly, the interactors reveal associations beyond one-carbon metabolism, covering also connections to nitrogen (N) metabolism, carbohydrate metabolism/photosynthesis, and proteostasis. Two of the interactions, one with the folate biosynthetic enzyme DIHYDROFOLATE REDUCTASE-THYMIDYLATE SYNTHASE 1 (AtDHFR-TS1) and another with N metabolism-associated glutamine synthetase 1;4 (AtGLN1;4), were further characterized. In silico and experimental analyses revealed G35/K36 and E330 as key residues for the binding of 5-F-THF in AtDHFR-TS1 and AtGLN1;4, respectively. Site-directed mutagenesis of AtGLN1;4 E330, which co-localizes with the ATP-binding pocket, abolished 5-F-THF binding as well as AtGLN1;4 activity. Furthermore, 5-F-THF was noted to competitively inhibit the activities of AtDHFR-TS1 and AtGLN1;4. In summary, we demonstrated a regulatory role for 5-F-THF in N metabolism, revealed 5-F-THF-mediated feedback regulation of folate biosynthesis, and identified a total of 14 previously unknown high-affinity binding cellular targets of 5-F-THF. Together, this sets a landmark toward understanding the role of folates in plant development.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021-10-11
2025
2025-11-17
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2117/446414
https://dx.doi.org/10.1093/plcell/koab198
url https://hdl.handle.net/2117/446414
https://dx.doi.org/10.1093/plcell/koab198
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:UPCommons. Portal del coneixement obert de la UPC
instname:Universitat Politècnica de Catalunya (UPC)
instname_str Universitat Politècnica de Catalunya (UPC)
reponame_str UPCommons. Portal del coneixement obert de la UPC
collection UPCommons. Portal del coneixement obert de la UPC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869408119818289152
score 15,811543

Similar Items