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Plant Physiol, April 2001, Vol. 125, pp. 2129-2138
Mobilization of Ca2+ by Cyclic ADP-Ribose from the
Endoplasmic Reticulum of Cauliflower
Florets1
Lorella
Navazio,2
Paola
Mariani, and
Dale
Sanders*
The Plant Laboratory, Department of Biology, University of York,
P.O. Box 373, York YO10 5YW, United Kingdom (L.N., D.S.); and
Dipartimento di Biologia, Università di Padova, Via U. Bassi
58/B, 35131 Padova, Italy (P.M.)
The NAD+ metabolite cADP-Rib (cADPR) elevates cytosolic
free Ca2+ in plants and thereby plays a central role in
signal transduction pathways evoked by the drought and stress hormone
abscisic acid. cADPR is known to mobilize Ca2+ from the
large vacuole of mature cells. To determine whether additional sites
for cADPR-gated Ca2+ release reside in plant cells,
microsomes from cauliflower (Brassica oleracea)
inflorescences were subfractionated on sucrose density gradients, and
the distribution of cADPR-elicited Ca2+ release was
monitored. cADPR-gated Ca2+ release was detected in the
heavy-density fractions associated with rough endoplasmic reticulum
(ER). cADPR-dependent Ca2+ release co-migrated with two ER
markers, calnexin and antimycin A-insensitive NADH-cytochrome
c reductase activity. To investigate the possibility
that contaminating plasma membrane in the ER-rich fractions was
responsible for the observed release, plasma membrane vesicles were
purified by aqueous two-phase partitioning, everted with Brij-58, and
loaded with Ca2+: These vesicles failed to respond to
cADPR. Ca2+ release evoked by cADPR at the ER was fully
inhibited by ruthenium red and 8-NH2-cADPR, a specific
antagonist of cADPR-gated Ca2+ release in animal cells. The
presence of a Ca2+ release pathway activated by cADPR at
higher plant ER reinforces the notion that, alongside the vacuole, the
ER participates in Ca2+ signaling.
1
This work was supported by the European
Molecular Biology Organization (award of a long-term fellowship to
L.N.), by the Ministero Università Ricerca Scientifica e
Tecnologica (to P.M.), and by the Biotechnology and Biological Sciences
Research Council (to D.S.).
2
Present address: Dipartimento di Biologia,
Università di Padova, Via U. Bassi 58/B, 35131 Padova, Italy.
*
Corresponding author: email ds10{at}york.ac.uk; fax
44-1904-434317.
© 2001 American Society of Plant Physiologists
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