Differential subplastidial localization and turnover of enzymes involved in isoprenoid biosynthesis in chloroplasts

Plastidial isoprenoids are a diverse group of metabolites with roles in photosynthesis, growth regulation, and interaction with the environment. The methylerythritol 4-phosphate (MEP) pathway produces the metabolic precursors of all types of plastidial isoprenoids. Proteomics studies in Arabidopsis thaliana have shown that all the enzymes of the MEP pathway are localized in the plastid stroma. However, immunoblot analysis of chloroplast subfractions showed that the first two enzymes of the pathway, deoxyxylulose 5-phosphate synthase (DXS) and reductoisomerase (DXR), can also be found in non-stromal fractions. Both transient and stable expression of GFP-tagged DXS and DXR proteins confirmed the presence of the fusion proteins in distinct subplastidial compartments. In particular, DXR-GFP was found to accumulate in relatively large vesicles that could eventually be released from chloroplasts, presumably to be degraded by an autophagy-independent process. Together, we propose that protein-specific mechanisms control the localization and turnover of the first two enzymes of the MEP pathway in Arabidopsis chloroplasts.This work was funded by grants from Generalitat de Catalunya (2014SGR-1434), Spanish Ministerio de Economia y Competitividad (BIO2014-59092-P and BIO2015-71703-REDT), and European Commission FP7 (TiMet, contract 245143) to MRC. CP and MOA received predoctoral fellowships from Spanish CSIC (JAE-PRE program) and MINECO (FPI program), respectively. EL is supported by a Mexican CONACYT predoctoral fellowship.Peer reviewe