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2.
Elife ; 82019 02 20.
Article in English | MEDLINE | ID: mdl-30785397

ABSTRACT

Pyrophosphate (PPi), a byproduct of macromolecule biosynthesis is maintained at low levels by soluble inorganic pyrophosphatases (sPPase) found in all eukaryotes. In plants, H+-pumping pyrophosphatases (H+-PPase) convert the substantial energy present in PPi into an electrochemical gradient. We show here, that both cold- and heat stress sensitivity of fugu5 mutants lacking the major H+-PPase isoform AVP1 is correlated with reduced SUMOylation. In addition, we show that increased PPi concentrations interfere with SUMOylation in yeast and we provide evidence that SUMO activating E1-enzymes are inhibited by micromolar concentrations of PPi in a non-competitive manner. Taken together, our results do not only provide a mechanistic explanation for the beneficial effects of AVP1 overexpression in plants but they also highlight PPi as an important integrator of metabolism and stress tolerance.


Subject(s)
Arabidopsis/physiology , Diphosphates/metabolism , Stress, Physiological , Sumoylation , Acclimatization , Arabidopsis/enzymology , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Cold Temperature , Hot Temperature , Inorganic Pyrophosphatase/metabolism , Isoenzymes/metabolism
3.
Plant Cell ; 27(12): 3383-96, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26589552

ABSTRACT

The presence of a large central vacuole is one of the hallmarks of a prototypical plant cell, and the multiple functions of this compartment require massive fluxes of molecules across its limiting membrane, the tonoplast. Transport is assumed to be energized by the membrane potential and the proton gradient established by the combined activity of two proton pumps, the vacuolar H(+)-pyrophosphatase (V-PPase) and the vacuolar H(+)-ATPase (V-ATPase). Exactly how labor is divided between these two enzymes has remained elusive. Here, we provide evidence using gain- and loss-of-function approaches that lack of the V-ATPase cannot be compensated for by increased V-PPase activity. Moreover, we show that increased V-ATPase activity during cold acclimation requires the presence of the V-PPase. Most importantly, we demonstrate that a mutant lacking both of these proton pumps is conditionally viable and retains significant vacuolar acidification, pointing to a so far undetected contribution of the trans-Golgi network/early endosome-localized V-ATPase to vacuolar pH.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Genome, Plant/genetics , Inorganic Pyrophosphatase/metabolism , Vacuolar Proton-Translocating ATPases/metabolism , Vacuoles/enzymology , Acclimatization , Arabidopsis/cytology , Arabidopsis/genetics , Arabidopsis/physiology , Arabidopsis Proteins/antagonists & inhibitors , Arabidopsis Proteins/genetics , Cold Temperature , Endosomes/enzymology , Flowers/cytology , Flowers/enzymology , Flowers/genetics , Flowers/physiology , Hydrogen-Ion Concentration , Inorganic Pyrophosphatase/antagonists & inhibitors , Inorganic Pyrophosphatase/genetics , Meristem/cytology , Meristem/enzymology , Meristem/genetics , Meristem/physiology , Mutagenesis, Insertional , Phenotype , Plant Roots/cytology , Plant Roots/enzymology , Plant Roots/genetics , Plant Roots/physiology , Seedlings/cytology , Seedlings/enzymology , Seedlings/genetics , Seedlings/physiology , Sequence Analysis, DNA , Vacuolar Proton-Translocating ATPases/antagonists & inhibitors , Vacuolar Proton-Translocating ATPases/genetics , trans-Golgi Network/enzymology
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