TY - JOUR
T1 - The flip side of the Arabidopsis type I proton-pumping pyrophosphatase (AVP1)
T2 - Using a transmembrane H gradient to synthesize pyrophosphate
AU - Scholz-Starke, Joachim
AU - Primo, Cecilia
AU - Yang, Jian
AU - Kandel, Raju
AU - Gaxiola, Roberto A.
AU - Hirschi, Kendal D.
N1 - Publisher Copyright:
© 2019 Scholz-Starke et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2019/1/25
Y1 - 2019/1/25
N2 - Energy partitioning and plant growth are mediated in part by a type I H-pumping pyrophosphatase (H-PPase). A canonical role for this transporter has been demonstrated at the tonoplast where it serves a job-sharing role with V-ATPase in vacuolar acidification. Here, we investigated whether the plant H-PPase from Arabidopsis also functions in “reverse mode” to synthesize PPi using the transmembrane H gradient. Using patch-clamp recordings on Arabidopsis vacuoles, we observed inward currents upon Pi application on the cytosolic side. These currents were strongly reduced in vacuoles from two independent H-PPase mutant lines (vhp1-1 and fugu5-1) lacking the classical PPi-induced outward currents related to H pumping, whereas they were significantly larger in vacuoles with engineered heightened expression of the H-PPase. Current amplitudes related to reverse-mode H transport depended on the membrane potential, cytosolic Pi concentration, and magnitude of the pH gradient across the tonoplast. Of note, experiments on vacuolar membrane- enriched vesicles isolated from yeast expressing the Arabidopsis H-PPase (AVP1) demonstrated Pi-dependent PPi synthase activity in the presence of a pH gradient. Our work establishes that a plant H-PPase can operate as a PPi synthase beyond its canonical role in vacuolar acidification and cytosolic PPi scavenging. We propose that the PPi synthase activity of H-PPase contributes to a cascade of events that energize plant growth.
AB - Energy partitioning and plant growth are mediated in part by a type I H-pumping pyrophosphatase (H-PPase). A canonical role for this transporter has been demonstrated at the tonoplast where it serves a job-sharing role with V-ATPase in vacuolar acidification. Here, we investigated whether the plant H-PPase from Arabidopsis also functions in “reverse mode” to synthesize PPi using the transmembrane H gradient. Using patch-clamp recordings on Arabidopsis vacuoles, we observed inward currents upon Pi application on the cytosolic side. These currents were strongly reduced in vacuoles from two independent H-PPase mutant lines (vhp1-1 and fugu5-1) lacking the classical PPi-induced outward currents related to H pumping, whereas they were significantly larger in vacuoles with engineered heightened expression of the H-PPase. Current amplitudes related to reverse-mode H transport depended on the membrane potential, cytosolic Pi concentration, and magnitude of the pH gradient across the tonoplast. Of note, experiments on vacuolar membrane- enriched vesicles isolated from yeast expressing the Arabidopsis H-PPase (AVP1) demonstrated Pi-dependent PPi synthase activity in the presence of a pH gradient. Our work establishes that a plant H-PPase can operate as a PPi synthase beyond its canonical role in vacuolar acidification and cytosolic PPi scavenging. We propose that the PPi synthase activity of H-PPase contributes to a cascade of events that energize plant growth.
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U2 - 10.1074/jbc.RA118.006315
DO - 10.1074/jbc.RA118.006315
M3 - Article
C2 - 30510138
AN - SCOPUS:85060659691
SN - 0021-9258
VL - 294
SP - 1290
EP - 1299
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 4
ER -