Acclimation of Photosynthesis to Changes in the Environment Results in Decreases of Oxidative Stress in Arabidopsis thaliana

The dynamic acclimation of photosynthesis plays an important role in increasing the fitness of a plant under variable light environments. Since acclimation is partially mediated by a glucose-6-phosphate/phosphate translocator 2 (GPT2), this study examined whether plants lacking GPT2, which consequently have defective acclimation to increases in light, are more susceptible to oxidative stress. To understand this mechanism, we used the model plant Arabidopsis thaliana [accession Wassilewskija-4 (Ws-4)] and compared it with mutants lacking GPT2. The plants were then grown at low light (LL) at 100 μmol m−2 s−1 for 7 weeks. For the acclimation experiments, a set of plants from LL was transferred to 400 μmol m−2 s−1 conditions for 7 days. Biochemical and physiological analyses showed that the gpt2 mutant plants had significantly greater activity for ascorbate peroxidase (APX), guiacol peroxidase (GPOX), and superoxide dismutase (SOD). Furthermore, the mutant plants had significantly lower maximum quantum yields of photosynthesis (Fv/Fm). A microarray analysis also showed that gpt2 plants exhibited a greater induction of stress-related genes relative to wild-type (WT) plants. We then concluded that photosynthetic acclimation to a higher intensity of light protects plants against oxidative stress.

Evidence for dual targeting of Arabidopsis plastidial glucose-6-phosphate transporter GPT1 to peroxisomes via the ER

ABSTRACTFormer studies on Arabidopsis glucose-6-phosphate/phosphate translocator isoforms GPT1 and GPT2 reported viability of gpt2 mutants, however an essential function for GPT1, manifesting as a variety of gpt1 defects in the heterozygous state during fertilization/seed set. Among other functions, GPT1 is important for pollen and embryo-sac development. Since previous work on enzymes of the oxidative pentose phosphate pathway (OPPP) revealed comparable effects, we investigated whether GPT1 might dually localize to plastids and peroxisomes. In reporter fusions, GPT2 was found at plastids, but GPT1 also at the endoplasmic reticulum (ER) and around peroxisomes. GPT1 contacted oxidoreductases and also peroxins that mediate import of peroxisomal membrane proteins from the ER, hinting at dual localization. Reconstitution in yeast proteoliposomes revealed that GPT1 preferentially exchanges glucose-6-phosphate for ribulose-5-phosphate. Complementation analyses of heterozygous gpt1 plants demonstrated that GPT2 is unable to compensate for GPT1 in plastids, whereas genomic GPT1 without transit peptide (enforcing ER/peroxisomal localization) increased gpt1 transmission significantly. Since OPPP activity in peroxisomes is essential during fertilization, and immuno-blot analyses hinted at unprocessed GPT1-specific bands, our findings suggest that GPT1 is indispensable at both plastids and peroxisomes. Together with the G6P-Ru5P exchange preference, dual targeting explains why GPT1 exerts functions distinct from GPT2 in Arabidopsis.One sentence summaryIn contrast to plastidial GPT2, GPT1 exhibits slightly different exchange preferences and alternatively targets the ER, from where the protein can be relocated to peroxisomes on demand.

Free energy landscape for the entire transport cycle of triose-phosphate/phosphate translocator

Secondary active transporters translocate their substrates using the electrochemical potentials of other chemicals, undergoing large-scale conformational changes. Despite extensive structural studies, the atomic details of the transport mechanism still remain elusive. Here we performed a series of all-atom molecular dynamics simulations of the triose-phosphate/phosphate translocator (TPT), which exports organic phosphates in the chloroplast stroma in strict counter exchange with inorganic phosphate (Pi). Biased sampling methods, including string method and umbrella sampling, successfully reproduced the conformational changes between the inward– and outward-facing states, along with the substrate binding. The free energy landscape of this entire TPT transition pathway demonstrated the alternating access and substrate translocation mechanisms, which revealed Pi is relayed by positively charged residues along the transition pathway. Furthermore, the conserved Glu207 functions as a “molecular switch”, linking the local substrate binding and the global conformational transition. Our results provide atomic-detailed insights into the energy coupling mechanism of antiporter.