Proton exchange by the vacuolar nitrate transporter CLCa is required for plant growth and nitrogen use efficiency.

Nitrate is a major nutrient and osmoticum for plants. To deal with fluctuating nitrate availability in soils, plants store this nutrient in their vacuoles. Chloride channel a (CLCa), a 2NO3-/1H+ exchanger localized to the vacuole in Arabidopsis (Arabidopsis thaliana), ensures this storage process.

Three-Dimensional Surface Rendering of ESCRT Proteins Microscopy Data Using UCSF Chimera Software.

Visualization of subcellular localization of ESCRT proteins and their interactions with different cellular compartments are critical to understand their function. This approach requires the generation of an important amount of 3D fluorescence microscopy data that is not always easy to visualize and apprehend.We describe a step-by-step protocol for 3D surface rendering of confocal microscopy acquisitions using the free software UCSF-Chimera, generating snapshots and animations to facilitate analysis and presentation of subcellular localization data.

Folding into an autophagosome: ATG5 sheds light on how plants do it.

Autophagosomes arise in yeast and animals from the sealing of a cup-shaped double-membrane precursor, the phagophore. The concerted action of about 30 evolutionarily conserved autophagy related (ATG) proteins lies at the core of this process. However, the mechanisms allowing phagophore generation and its differentiation into a sealed autophagosome are still not clear in detail, and very little is known in plants.

ATG5 defines a phagophore domain connected to the endoplasmic reticulum during autophagosome formation in plants.

Autophagosomes are the organelles responsible for macroautophagy and arise, in yeast and animals, from the sealing of a cup-shaped double-membrane precursor, the phagophore. How the phagophore is generated and grows into a sealed autophagosome is still not clear in detail, and unknown in plants. This is due, in part, to the scarcity of structurally informative, real-time imaging data of the required protein machinery at the phagophore formation site.

The RNA binding protein Tudor-SN is essential for stress tolerance and stabilizes levels of stress-responsive mRNAs encoding secreted proteins in Arabidopsis.

Tudor-SN (TSN) copurifies with the RNA-induced silencing complex in animal cells where, among other functions, it is thought to act on mRNA stability via the degradation of specific dsRNA templates. In plants, TSN has been identified biochemically as a cytoskeleton-associated RNA binding activity. In eukaryotes, it has recently been identified as a conserved primary target of programmed cell death-associated proteolysis.

Functional studies of shaggy/glycogen synthase kinase 3 phosphorylation sites in Drosophila melanogaster.

Early studies of glycogen synthase kinase 3 (GSK-3) in mammalian systems focused on its pivotal role in glycogen metabolism and insulin-mediated signaling. It is now recognized that GSK-3 is central to a number of diverse signaling systems. Here, we show that the major form of the kinase Shaggy (Sgg), the GSK-3 fly ortholog, is negatively regulated during insulin-like/phosphatidylinositol 3-kinase (PI3K) signaling in vivo.

Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase.

Glutathione transferases (GST; EC 2.5.1.

Identification of proteins regulated by cross-talk between drought and hormone pathways in Arabidopsis wild-type and auxin-insensitive mutants, axr1 and axr2.

Ten proteins differentially regulated by progressive drought stress in Arabidopsis Columbia wild-type, axr1-3 and axr2-1auxin-insensitive mutants, were identified from internal amino acid microsequencing. These proteins fell into two categories: (i) stress-related proteins, known to be induced by rapid water stress via abscisic acid (ABA)-dependent or -independent pathways [late embryogenesis abundant (LEA)-like and heat shock cognate (HS) 70, respectively], or in response to pathogens or oxidative stress [β-1,3 glucanase (BG), annexin] and (ii) metabolic enzymes [glutamine synthetase (GS), fructokinase (Frk), caffeoyl-CoA-3-O-methyltransferase (CCoAOMT)]. The differential behaviour of these proteins highlighted a role for AXR2 and/or AXR1 in the regulation of their abundance during drought adaptation.