Manganese （Mn） is an essential catalytic metal in the Mn-cluster that oxidizes water to produce oxygen dur- ing photosynthesis. However, the transport protein（s） responsible for Mn2＋ import into the chloroplast r...Manganese （Mn） is an essential catalytic metal in the Mn-cluster that oxidizes water to produce oxygen dur- ing photosynthesis. However, the transport protein（s） responsible for Mn2＋ import into the chloroplast re- mains unknown. Here, we report the characterization ofArabidopsis CMT1 （Chloroplast Manganese Trans- porter 1）, an evolutionarily conserved protein in the Uncharacterized Protein Family 0016 （UPFO016）, that is required for manganese accumulation into the chloroplast. CMT1 is expressed primarily in green tissues, and its encoded product is localized in the inner envelope membrane of the chloroplast. Disruption of CMT1 in the T-DNA insertional mutant cmtl-1 resulted in stunted plant growth, defective thylakoid stacking, and severe reduction of photosystem II complexes and photosynthetic activity. Consistent with reduced oxy- gen evolution capacity, the mutant chloroplasts contained less manganese than the wild-type ones. In sup- port of its function as a Mn transporter, CMT1 protein supported the growth and enabled Mn2＋ accumula- tion in the yeast cells of Mn2＋-uptake deficient mutant （3smfl）. Taken together, our results indicate that CMT1 functions as an inner envelope Mn transporter responsible for chloroplast Mn2＋ uptake.展开更多
Excess phosphate(Pi)is stored into the vacuole through Pi transporters so that cytoplasmic Pi levels remain stable in plant cells.We hypothesized that the vacuolar Pi transporters may harbor a Pi-sensing mechanism so ...Excess phosphate(Pi)is stored into the vacuole through Pi transporters so that cytoplasmic Pi levels remain stable in plant cells.We hypothesized that the vacuolar Pi transporters may harbor a Pi-sensing mechanism so that they are activated to deliver Pi into the vacuole only when cytosolic Pi reaches a threshold high level.We tested this hypothesis using Vacuolar Phosphate Transporter 1(VPT1),a SPX domain-containing vacuolar Pi transporter,as a model.Recent studies have defined SPX as a Pi-sensing module that binds inositol polyphosphate signaling molecules(InsPs)produced at high cellular Pi status.We showed here that Pi-deficient conditions or mutation of the SPX domain severely impaired the transport activity of VPT1.We further identified an auto-inhibitory domain in VPT1 that suppresses its transport activity.Taking together the results from detailed structure-function analyses,our study suggests that VPT1 is in the auto-inhibitory state when Pi status is low,whereas at high cellular Pi status InsPs are produced and bind SPX domain to switch on VPT1 activity to deliver Pi into the vacuole.This thus provides an auto-regulatory mechanism for VPT1-mediated Pi sensing and homeostasis in plant cells.展开更多
Chlorophyll(Chl)is essential for photosynthetic reactions and chloroplast development.While the enzymatic pathway for Chl biosynthesis is well established,the regulatory mechanism underlying the homeostasis of Chl lev...Chlorophyll(Chl)is essential for photosynthetic reactions and chloroplast development.While the enzymatic pathway for Chl biosynthesis is well established,the regulatory mechanism underlying the homeostasis of Chl levels remains largely unknown.In this study,we identified CBD1(Chlorophyll Biosynthetic Defect1),which functions in the regulation of chlorophyll biosynthesis.The CBD1 gene was expressed specifically in green tissues and its protein product was embedded in the thylakoid membrane.Furthermore,CBD1 was precisely co-expressed and functionally correlated with GUN5(Genome Uncoupled 5).Analysis of chlorophyll metabolic intermediates indicated that cbd1 and cbd1gun5 mutants over-accumulatedmagnesium protoporphyrin Ⅸ(Mg-Proto Ⅸ).In addition,the cbd1 mutant thylakoid contained less Mg than the wild type not only as a result of lower Chl content,but also implicating CBD1 in Mg transport.This was supported by the finding that CBD1 complemented a Mg^(2+)uptake-deficient Salmonella strain under low Mg conditions.Taken together,these results indicate that CBD1 functions synergistically with CHLH/GUN5 in Mg-Proto IX processing,and may serve as a Mg-transport protein to maintain Mg homeostasis in the chloroplast.展开更多
文摘Manganese （Mn） is an essential catalytic metal in the Mn-cluster that oxidizes water to produce oxygen dur- ing photosynthesis. However, the transport protein（s） responsible for Mn2＋ import into the chloroplast re- mains unknown. Here, we report the characterization ofArabidopsis CMT1 （Chloroplast Manganese Trans- porter 1）, an evolutionarily conserved protein in the Uncharacterized Protein Family 0016 （UPFO016）, that is required for manganese accumulation into the chloroplast. CMT1 is expressed primarily in green tissues, and its encoded product is localized in the inner envelope membrane of the chloroplast. Disruption of CMT1 in the T-DNA insertional mutant cmtl-1 resulted in stunted plant growth, defective thylakoid stacking, and severe reduction of photosystem II complexes and photosynthetic activity. Consistent with reduced oxy- gen evolution capacity, the mutant chloroplasts contained less manganese than the wild-type ones. In sup- port of its function as a Mn transporter, CMT1 protein supported the growth and enabled Mn2＋ accumula- tion in the yeast cells of Mn2＋-uptake deficient mutant （3smfl）. Taken together, our results indicate that CMT1 functions as an inner envelope Mn transporter responsible for chloroplast Mn2＋ uptake.
基金the China Postdoctoral Science Foundation 2020M683537(to M.L)National Natural Science Foundation of China 32200216(to M.L)the National Science Foundation MCB 2041585(to S.L).
文摘Excess phosphate(Pi)is stored into the vacuole through Pi transporters so that cytoplasmic Pi levels remain stable in plant cells.We hypothesized that the vacuolar Pi transporters may harbor a Pi-sensing mechanism so that they are activated to deliver Pi into the vacuole only when cytosolic Pi reaches a threshold high level.We tested this hypothesis using Vacuolar Phosphate Transporter 1(VPT1),a SPX domain-containing vacuolar Pi transporter,as a model.Recent studies have defined SPX as a Pi-sensing module that binds inositol polyphosphate signaling molecules(InsPs)produced at high cellular Pi status.We showed here that Pi-deficient conditions or mutation of the SPX domain severely impaired the transport activity of VPT1.We further identified an auto-inhibitory domain in VPT1 that suppresses its transport activity.Taking together the results from detailed structure-function analyses,our study suggests that VPT1 is in the auto-inhibitory state when Pi status is low,whereas at high cellular Pi status InsPs are produced and bind SPX domain to switch on VPT1 activity to deliver Pi into the vacuole.This thus provides an auto-regulatory mechanism for VPT1-mediated Pi sensing and homeostasis in plant cells.
基金supported by the National Natural Science Foundation of China(grant no.31900220 to C.Z.,and 31770267 to W.L.)the National Science Foundation(MCB-1714795 to S.L.).
文摘Chlorophyll(Chl)is essential for photosynthetic reactions and chloroplast development.While the enzymatic pathway for Chl biosynthesis is well established,the regulatory mechanism underlying the homeostasis of Chl levels remains largely unknown.In this study,we identified CBD1(Chlorophyll Biosynthetic Defect1),which functions in the regulation of chlorophyll biosynthesis.The CBD1 gene was expressed specifically in green tissues and its protein product was embedded in the thylakoid membrane.Furthermore,CBD1 was precisely co-expressed and functionally correlated with GUN5(Genome Uncoupled 5).Analysis of chlorophyll metabolic intermediates indicated that cbd1 and cbd1gun5 mutants over-accumulatedmagnesium protoporphyrin Ⅸ(Mg-Proto Ⅸ).In addition,the cbd1 mutant thylakoid contained less Mg than the wild type not only as a result of lower Chl content,but also implicating CBD1 in Mg transport.This was supported by the finding that CBD1 complemented a Mg^(2+)uptake-deficient Salmonella strain under low Mg conditions.Taken together,these results indicate that CBD1 functions synergistically with CHLH/GUN5 in Mg-Proto IX processing,and may serve as a Mg-transport protein to maintain Mg homeostasis in the chloroplast.
