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

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

DOI：
10.1093/plcell/koab198
复制
发表时间：
2021
期刊：
The Plant Cell
影响因子：
--
通讯作者：
Youli Xiao
Youli Xiao
中科院分区：
--
文献类型：
--
作者：
Weichao Li;Qiuju Liang;Ratnesh Chandra Mishra;Raul Sanchez-Munoz;Huan Wang;Xin Chen;Dominique Van Der Straeten;Chunyi Zhang;Youli Xiao

文献摘要

相似文献

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.