scholarly journals Ubiquitin-dependent chloroplast-associated protein degradation in plants

Science ◽  
2019 ◽  
Vol 363 (6429) ◽  
pp. eaav4467 ◽  
Author(s):  
Qihua Ling ◽  
William Broad ◽  
Raphael Trösch ◽  
Mats Töpel ◽  
Tijen Demiral Sert ◽  
...  
Keyword(s):  
Protein Degradation ◽  
Forward Genetics ◽  
26S Proteasome ◽  
Chloroplast Envelope ◽  
Outer Envelope ◽  
Aaa Atpase ◽  
Ubiquitin E3 Ligase ◽  
Outer Envelope Membrane ◽  
Underlying Mechanisms ◽  
Envelope Membranes

Chloroplasts contain thousands of nucleus-encoded proteins that are imported from the cytosol by translocases in the chloroplast envelope membranes. Proteolytic regulation of the translocases is critically important, but little is known about the underlying mechanisms. We applied forward genetics and proteomics in Arabidopsis to identify factors required for chloroplast outer envelope membrane (OEM) protein degradation. We identified SP2, an Omp85-type β-barrel channel of the OEM, and CDC48, a cytosolic AAA+ (ATPase associated with diverse cellular activities) chaperone. Both proteins acted in the same pathway as the ubiquitin E3 ligase SP1, which regulates OEM translocase components. SP2 and CDC48 cooperated to bring about retrotranslocation of ubiquitinated substrates from the OEM (fulfilling conductance and motor functions, respectively), enabling degradation of the substrates by the 26S proteasome in the cytosol. Such chloroplast-associated protein degradation (CHLORAD) is vital for organellar functions and plant development.

Mechanism of protein import across the chloroplast envelope

2000 ◽  
Vol 28 (4) ◽  
pp. 485-491 ◽  
Author(s):  
K. Chen ◽  
X. Chen ◽  
D. J. Schnell
Keyword(s):  
Protein Import ◽  
Essential Elements ◽  
Chloroplast Envelope ◽  
Envelope Membrane ◽  
Outer Envelope ◽  
Double Membrane ◽  
Protein Subunits ◽  
Targeting Signals ◽  
Outer Envelope Membrane ◽  
Envelope Membranes

The development and maintenance of chloroplasts relies on the contribution of protein subunits from both plastid and nuclear genomes. Most chloroplast proteins are encoded by nuclear genes and are post-translationally imported into the organelle across the double membrane of the chloroplast envelope. Protein import into the chloroplast consists of two essential elements: the specific recognition of the targeting signals (transit sequences) of cytoplasmic preproteins by receptors at the outer envelope membrane and the subsequent translocation of preproteins simultaneously across the double membrane of the envelope. These processes are mediated via the co-ordinate action of protein translocon complexes in the outer (Toe apparatus) and inner (Tic apparatus) envelope membranes.

scholarly journals Synthesis and transfer of galactolipids in the chloroplast envelope membranes of Arabidopsis thaliana

2016 ◽  
Vol 113 (38) ◽  
pp. 10714-10719 ◽  
Author(s):  
Amélie A. Kelly ◽  
Barbara Kalisch ◽  
Georg Hölzl ◽  
Sandra Schulze ◽  
Juliane Thiele ◽  
...  
Keyword(s):  
Fusion Proteins ◽  
Chloroplast Envelope ◽  
Envelope Membrane ◽  
Lipid Transfer ◽  
Terminal Sequence ◽  
Outer Envelope ◽  
Outer Envelope Membrane ◽  
Envelope Reconstruction ◽  
Envelope Membranes

Galactolipids [monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)] are the hallmark lipids of photosynthetic membranes. The galactolipid synthases MGD1 and DGD1 catalyze consecutive galactosyltransfer reactions but localize to the inner and outer chloroplast envelopes, respectively, necessitating intermembrane lipid transfer. Here we show that the N-terminal sequence of DGD1 (NDGD1) is required for galactolipid transfer between the envelopes. Different diglycosyllipid synthases (DGD1, DGD2, and Chloroflexus glucosyltransferase) were introduced into the dgd1-1 mutant of Arabidopsis in fusion with N-terminal extensions (NDGD1 and NDGD2) targeting to the outer envelope. Reconstruction of DGDG synthesis in the outer envelope membrane was observed only with diglycosyllipid synthase fusion proteins carrying NDGD1, indicating that NDGD1 enables galactolipid translocation between envelopes. NDGD1 binds to phosphatidic acid (PA) in membranes and mediates PA-dependent membrane fusion in vitro. These findings provide a mechanism for the sorting and selective channeling of lipid precursors between the galactolipid pools of the two envelope membranes.

scholarly journals A new chloroplast protein import intermediate reveals distinct translocation machineries in the two envelope membranes: energetics and mechanistic implications.

1996 ◽  
Vol 132 (1) ◽  
pp. 63-75 ◽  
Author(s):  
S V Scott ◽  
S M Theg
Keyword(s):  
Membrane Transport ◽  
Protein Import ◽  
Chloroplast Envelope ◽  
Envelope Membrane ◽  
Outer Envelope ◽  
Stromal Compartment ◽  
Chloroplast Protein Import ◽  
Chloroplast Protein ◽  
Outer Envelope Membrane ◽  
Envelope Membranes

Chloroplast protein import presents a complex membrane traversal problem: precursor proteins must cross two envelope membranes to reach the stromal compartment. This work characterizes a new chloroplast protein import intermediate which has completely traversed the outer envelope membrane but has not yet reached the stroma. The existence of this intermediate demonstrates that distinct protein transport machineries are present in both envelope membranes, and that they are able to operate independently of one another under certain conditions. Energetic characterization of this pathway led to the identification of three independent energy-requiring steps: binding of the precursor to the outer envelope membrane, outer membrane transport, and inner membrane transport. Localization of the sites of energy utilization for each of these steps, as well as their respective nucleotide specificities, suggest that three different ATPases mediate chloroplast envelope transport.

scholarly journals AAA+ ATPases in Protein Degradation: Structures, Functions and Mechanisms

Biomolecules ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 629 ◽  
Author(s):  
Shuwen Zhang ◽  
Youdong Mao
Keyword(s):  
Protein Degradation ◽  
Conformational Changes ◽  
Dynamic Models ◽  
Three Dimensional ◽  
26S Proteasome ◽  
Aaa Atpase ◽  
Substrate Protein ◽  
Function Relationship ◽  
Atp Binding And Hydrolysis ◽  
Aaa Atpases

Adenosine triphosphatases (ATPases) associated with a variety of cellular activities (AAA+), the hexameric ring-shaped motor complexes located in all ATP-driven proteolytic machines, are involved in many cellular processes. Powered by cycles of ATP binding and hydrolysis, conformational changes in AAA+ ATPases can generate mechanical work that unfolds a substrate protein inside the central axial channel of ATPase ring for degradation. Three-dimensional visualizations of several AAA+ ATPase complexes in the act of substrate processing for protein degradation have been resolved at the atomic level thanks to recent technical advances in cryogenic electron microscopy (cryo-EM). Here, we summarize the resulting advances in structural and biochemical studies of AAA+ proteases in the process of proteolysis reactions, with an emphasis on cryo-EM structural analyses of the 26S proteasome, Cdc48/p97 and FtsH-like mitochondrial proteases. These studies reveal three highly conserved patterns in the structure–function relationship of AAA+ ATPase hexamers that were observed in the human 26S proteasome, thus suggesting common dynamic models of mechanochemical coupling during force generation and substrate translocation.

scholarly journals Insertion of plastidic β-barrel proteins into the outer envelopes of plastids involves an intermembrane space intermediate formed with Toc75-V/OEP80

2021 ◽  
Author(s):  
Lucia E Gross ◽  
Anna Klinger ◽  
Nicole Spies ◽  
Theresa Ernst ◽  
Nadine Flinner ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Protein Import ◽  
Membrane Insertion ◽  
Intermembrane Space ◽  
Envelope Membrane ◽  
Outer Envelope ◽  
Pea Proteins ◽  
Outer Envelope Membrane ◽  
Correct Topology ◽  
Shed Light

Abstract The insertion of organellar membrane proteins with the correct topology requires the following: First, the proteins must contain topogenic signals for translocation across and insertion into the membrane. Second, proteinaceous complexes in the cytoplasm, membrane, and lumen of organelles are required to drive this process. Many complexes required for the intracellular distribution of membrane proteins have been described, but the signals and components required for the insertion of plastidic β-barrel-type proteins into the outer membrane are largely unknown. The discovery of common principles is difficult, as only a few plastidic β-barrel proteins exist. Here, we provide evidence that the plastidic outer envelope β-barrel proteins OEP21, OEP24, and OEP37 from pea (Pisum sativum) and Arabidopsis thaliana contain information defining the topology of the protein. The information required for translocation of pea proteins across the outer envelope membrane is present within the six N-terminal β-strands. This process requires the action of TOC (translocon of the outer chloroplast membrane). After translocation into the intermembrane space, β-barrel proteins interact with TOC75-V, as exemplified by OEP37 and P39, and are integrated into the membrane. The membrane insertion of plastidic β-barrel proteins is affected by mutation of the last β-strand, suggesting that this strand contributes to the insertion signal. These findings shed light on the elements and complexes involved in plastidic β-barrel protein import.

Nitrate metabolism in relation to the aspartate-type C-4 pathway of photosynthesis in Eleusine coracana

1974 ◽  
Vol 52 (12) ◽  
pp. 2599-2605 ◽  
Author(s):  
C. K. M. Rathnam ◽  
V. S. R. Das
Keyword(s):  
Glutamate Dehydrogenase ◽  
Eleusine Coracana ◽  
Bundle Sheath ◽  
Chloroplast Envelope ◽  
Mesophyll Cells ◽  
Bundle Sheath Cells ◽  
Nitrate Metabolism ◽  
Type C ◽  
Envelope Membranes ◽  
Sheath Cells

The intercellular and intracellular distributions of nitrate assimilating enzymes were studied. Nitrate reductase was found to be localized on the chloroplast envelope membranes. The chloroplastic NADPH – glutamate dehydrogenase was concentrated in the mesophyll cells. The extrachloroplastic NADH – glutamate dehydrogenase was localized in the bundle sheath cells. Glutamate synthesized in the mesophyll chloroplasts was interpreted to be utilized exclusively in the synthesis of aspartate, while in the bundle sheath cells it was thought to be consumed in other cellular metabolic processes. Based on the results, a scheme is proposed to account for the nitrate metabolism in the leaves of Eleusine coracana Gaertn. in relation to its aspartate-type C-4 pathway of photosynthesis.

scholarly journals Ccr4 is a novel shuttle factor required for ubiquitin-dependent proteolysis by the 26S proteasome

2020 ◽  
Author(s):  
Ganapathi Kandasamy ◽  
Ashis Kumar Pradhan ◽  
R Palanimurugan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Degradation ◽  
Ubiquitin Proteasome System ◽  
Proteasomal Degradation ◽  
Rna Degradation ◽  
26S Proteasome ◽  
Cellular Proteins ◽  
Cellular Homeostasis ◽  
Substrate Degradation ◽  
Ubiquitin Proteasome

AbstractDegradation of short-lived and abnormal proteins are essential for normal cellular homeostasis. In eukaryotes, such unstable cellular proteins are selectively degraded by the ubiquitin proteasome system (UPS). Furthermore, abnormalities in protein degradation by the UPS have been linked to several human diseases. Ccr4 protein is a known component of the Ccr4-Not complex, which has established roles in transcription, mRNA de-adenylation and RNA degradation etc. Excitingly in this study, we show that Ccr4 protein has a novel function as a shuttle factor that promotes ubiquitin-dependent degradation of short-lived proteins by the 26S proteasome. Using a substrate of the well-studied ubiquitin fusion degradation (UFD) pathway, we found that its UPS-mediated degradation was severely impaired upon deletion of CCR4 in Saccharomyces cerevisiae. Additionally, we show that Ccr4 binds to cellular ubiquitin conjugates and the proteasome. In contrast to Ccr4, most other subunits of the Ccr4-Not complex proteins are dispensable for UFD substrate degradation. From our findings we conclude that Ccr4 functions in the UPS as a shuttle factor targeting ubiquitylated substrates for proteasomal degradation.

scholarly journals TIP family aquaporins play role in chloroplast osmoregulation and photosynthesis

2020 ◽  
Author(s):  
Azeez Beebo ◽  
Ahmad Zia ◽  
Christopher R. Kinzel ◽  
Andrei Herdean ◽  
Karim Bouhidel ◽  
...  
Keyword(s):  
Electron Transport ◽  
Oxygen Evolution ◽  
Photosynthetic Electron Transport ◽  
Chloroplast Envelope ◽  
Photosynthetic Oxygen Evolution ◽  
Wild Type ◽  
Thylakoid Lumen ◽  
Photosynthetic Oxygen ◽  
Osmotic Treatment ◽  
Envelope Membranes

SUMMARYPhotosynthetic oxygen evolution by photosystem II requires water supply into the chloroplast to reach the thylakoid lumen. A rapid water flow is also required into the chloroplast for optimal oxygen evolution and to overcome osmotic stress. The mechanisms governing water transport in chloroplasts are largely unexplored. Previous proteomics indicated the presence of three aquaporins from the tonoplast intrinsic protein (TIP) family, TIP1;1, TIP1;2 and TIP2;1, in chloroplast membranes of Arabidopsis thaliana. Here we revisited their location and studied their role in chloroplasts. Localization experiments indicated that TIP2;1 resides in the thylakoid, whereas TIP1;2 is present in both thylakoid and envelope membranes. Mutants lacking TIP1;2 and/or TIP2;1 did not display a macroscopic phenotype when grown under standard conditions. The mutant chloroplasts and thylakoids underwent less volume changes than the corresponding wild type preparations upon osmotic treatment and in the light. Significantly reduced rates of photosynthetic electron transport were obtained in the mutant leaves, with implications on the CO2 fixation rates. However, electron transport rates did not significantly differ between mutants and wild type when isolated thylakoids were examined. Less acidification of the thylakoid lumen was measured in mutants thylakoids, resulting in a slower induction of delta pH-dependent photoprotective mechanisms. These results identify TIP1;2 and TIP2;1 as chloroplast proteins and highlight their importance for osmoregulation and optimal photosynthesis. A third aquaporin, TIP1;1, is present in the chloroplast envelope, and may play role in photosynthesis under excessive light conditions, as based on the weak photosynthetic phenotype of its mutant.

scholarly journals Adaptive mitochondrial regulation of the proteasome

2020 ◽  
Author(s):  
Thomas Meul ◽  
Korbinian Berschneider ◽  
Sabine Schmitt ◽  
Christoph H. Mayr ◽  
Laura F. Mattner ◽  
...  
Keyword(s):  
Protein Degradation ◽  
Transcriptional Activation ◽  
Krebs Cycle ◽  
Mitochondrial Metabolism ◽  
26S Proteasome ◽  
Metabolic Reprogramming ◽  
Metabolic Adaptation ◽  
Respiratory Dysfunction ◽  
Therapeutic Implications ◽  
Mtorc1 Pathway

SummaryThe proteasome is the main proteolytic system for targeted protein degradation in the cell. Its function is fine-tuned according to cellular needs. Regulation of proteasome function by mitochondrial metabolism, however, is unknown.Here, we demonstrate that mitochondrial dysfunction reduces the assembly and activity of the 26S proteasome in the absence of oxidative stress. Impaired respiratory complex I function leads to metabolic reprogramming of the Krebs cycle and deficiency in aspartate. Aspartate supplementation activates assembly and activity of 26S proteasomes via transcriptional activation of the proteasome assembly factors p28 and Rpn6. This metabolic adaptation of 26S proteasome function involves sensing of aspartate via the mTORC1 pathway. Metformin treatment of primary human cells similarly reduced assembly and activity of 26S proteasome complexes, which was fully reversible and rescued by supplementation of aspartate or pyruvate. Of note, respiratory dysfunction conferred resistance towards the proteasome inhibitor Bortezomib.Our study uncovers a fundamental novel mechanism of how mitochondrial metabolism adaptively adjusts protein degradation by the proteasome. It thus unravels unexpected consequences of defective mitochondrial metabolism in disease or drug-targeted mitochondrial reprogramming for proteasomal protein degradation in the cell. As metabolic inhibition of proteasome function can be alleviated by treatment with aspartate or pyruvate, our results also have therapeutic implications.

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