000127019 001__ 127019
000127019 005__ 20240319081031.0
000127019 0247_ $$2doi$$a10.1016/j.xplc.2022.100512
000127019 0248_ $$2sideral$$a134393
000127019 037__ $$aART-2022-134393
000127019 041__ $$aeng
000127019 100__ $$aDi, X.
000127019 245__ $$aMEP pathway products allosterically promote monomerization of deoxy-D-xylulose-5-phosphate synthase to feedback-regulate their supply
000127019 260__ $$c2022
000127019 5060_ $$aAccess copy available to the general public$$fUnrestricted
000127019 5203_ $$aIsoprenoids are a very large and diverse family of metabolites required by all living organisms. All isoprenoids derive from the double-bond isomers isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are produced by the methylerythritol 4-phosphate (MEP) pathway in bacteria and plant plastids. It has been reported that IPP and DMAPP feedback-regulate the activity of deoxyxylulose 5-phosphate synthase (DXS), a dimeric enzyme that catalyzes the main flux-controlling step of the MEP pathway. Here we provide experimental insights into the underlying mechanism. Isothermal titration calorimetry and dynamic light scattering approaches showed that IPP and DMAPP can allosterically bind to DXS in vitro, causing a size shift. In silico ligand binding site analysis and docking calculations identified a potential allosteric site in the contact region between the two monomers of the active DXS dimer. Modulation of IPP and DMAPP contents in vivo followed by immunoblot analyses confirmed that high IPP/DMAPP levels resulted in monomerization and eventual aggregation of the enzyme in bacterial and plant cells. Loss of the enzymatically active dimeric conformation allows a fast and reversible reduction of DXS activity in response to a sudden increase or decrease in IPP/DMAPP supply, whereas aggregation and subsequent removal of monomers that would otherwise be available for dimerization appears to be a more drastic response in the case of persistent IPP/DMAPP overabundance (e.g., by a blockage in their conversion to downstream isoprenoids). Our results represent an important step toward understanding the regulation of the MEP pathway and rational design of biotechnological endeavors aimed at increasing isoprenoid contents in microbial and plant systems.
000127019 536__ $$9info:eu-repo/grantAgreement/ES/AEI/PCI2021-121941$$9info:eu-repo/grantAgreement/ES/AEI/PID2020-115810GB-I00$$9info:eu-repo/grantAgreement/ES/CSIC/PIE-202040E299$$9info:eu-repo/grantAgreement/EC/FP7/627639/EU/Revealing the hidden secrets of the MEP pathway to engineer new bio-resources for humanity/BIOHELP$$9info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/BES-2017-080739$$9info:eu-repo/grantAgreement/ES/MINECO/FEDER/BFU2016-78232-P
000127019 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000127019 590__ $$a10.5$$b2022
000127019 591__ $$aPLANT SCIENCES$$b8 / 239 = 0.033$$c2022$$dQ1$$eT1
000127019 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b27 / 285 = 0.095$$c2022$$dQ1$$eT1
000127019 592__ $$a2.985$$b2022
000127019 593__ $$aBiochemistry$$c2022$$dQ1
000127019 593__ $$aBiotechnology$$c2022$$dQ1
000127019 593__ $$aPlant Science$$c2022$$dQ1
000127019 593__ $$aMolecular Biology$$c2022$$dQ1
000127019 593__ $$aCell Biology$$c2022$$dQ1
000127019 594__ $$a12.7$$b2022
000127019 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000127019 700__ $$0(orcid)0000-0003-1885-4365$$aOrtega-Alarcon, David$$uUniversidad de Zaragoza
000127019 700__ $$aKakumanu, Ramu
000127019 700__ $$aIglesias-Fernandez, Javier
000127019 700__ $$aDiaz, Lucia
000127019 700__ $$aBaidoo, Edward E.K.
000127019 700__ $$0(orcid)0000-0001-5702-4538$$aVelazquez-Campoy, Adrian$$uUniversidad de Zaragoza
000127019 700__ $$aRodríguez-Concepción, Manuel
000127019 700__ $$aPerez-Gil, Jordi
000127019 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000127019 773__ $$g4, 3 (2022), 100512 [12 pp.]$$tPlant Communications$$x2590-3462
000127019 8564_ $$s1802360$$uhttps://zaguan.unizar.es/record/127019/files/texto_completo.pdf$$yVersión publicada
000127019 8564_ $$s2788409$$uhttps://zaguan.unizar.es/record/127019/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000127019 909CO $$ooai:zaguan.unizar.es:127019$$particulos$$pdriver
000127019 951__ $$a2024-03-18-17:09:49
000127019 980__ $$aARTICLE