Professional Documents
Culture Documents
Kock Susy Invertases
Kock Susy Invertases
UDP
UDPG
VIN
VPEc
Addresses
Plant Molecular and Cellular Biology Program, Horticultural Sciences
Department, University of Florida, Gainesville, Florida 32611, USA
e-mail: kek@ifas.UFL.edu
Figure 1
uridine di-phosphate
UDPglucose
vacuolar invertase
vacuolar processing enzyme g
Introduction
The only known enzymatic paths of sucrose cleavage in
plants are catalyzed by invertases (sucrose H2O !
glucose fructose) and sucrose synthases (sucrose
UDP ! fructose UDPglucose) (Figure 1). Both of
these paths typically degrade sucrose in vivo but the
products of their reactions differ in important ways
[1,2]. Invertases produce glucose instead of UDPglucose
(UDPG), and thus also form twice as many hexoses.
Either of these features could give invertases a greater
capacity to stimulate specific sugar sensors [39]. The
resulting signals can alter expression of diverse genes
[3,10,11], so invertases can potentially be strong effectors
of widely varying processes. These include the biosynthesis and perception of hormones such as abscisic acid
(ABA). In addition, both sugar and hormone signals can
also affect the expression of the genes that encode sucrose
synthase and invertase [13,5,7,12,13,14,15,16,17]. This
review explores the hypothesis that sucrose metabolism
lies at the heart of a sensitive, self-regulatory developmental system in plants. Its influence appears to be
balanced by the capacity for sensing sucrose itself,
Hexose signals
Glucose
Invertase
Fructose
Hexose
signals
Sucrose
Sucrose synthase
Fructose
UDP glucose
Sucrose signals
Current Opinion in Plant Biology
DOI 10.1016/j.pbi.2004.03.014
Abbreviations
ABA
abscisic acid
CIN
cytoplasmic invertase
CWIN cell wall invertase
ENOD early nodulation
PPi
inorganic pyrophosphate
PPV
precursor protease vesicle
SnRK SNF-related kinase
SUS
sucrose synthase
www.sciencedirect.com
Phloem
Sucrose
Cell wall
space
Plasmodesmata
Sucrose
CIN
G+F
G+F
SUS
UDPG + F
VIN
G+F
The physical path of sucrose movement and site of its cleavage are
central not only to mechanisms of import but also to the sugar signals
generated. The mechanisms that regulate sucrose movement and
cleavage also differ for each compartment. Depending on the path of
sucrose entry into an importing structure (i.e. via plasmodesmata or
across the cell wall space), sucrose can be cleaved by cell wall
invertase (CWIN), cytoplasmic invertase (CIN), sucrose synthase (SUS),
or vacuolar invertase (VIN). Cytoplasmic sucrose cleavage typically
produces limited hexoses or hexose-based sugar signals because
of the generally low activity of CIN and the production of UDPG instead
of glucose by SUS. In contrast, sucrose that enters sinks via the cell
wall space can generate significant amounts of glucose and other
hexoses if CWIN is active. Glucose and fructose can, in turn, initiate
hexose-based signals both at the membrane and during subsequent
metabolism in the cytoplasm. Similarly, abundant hexoses and
hexose-based signals can be generated in the vacuole by VIN; this
constitutes the primary site of sucrose cleavage during the expansion
phase of most sink tissues. F, fructose; G, glucose.
Current Opinion in Plant Biology 2004, 7:235246
deposition [17,3840]). Overall, however, diverse evidence supports the contention that the balance between
invertase and sucrose synthase activity can alter plant
development through differential effects on sugar signaling systems.
Relative contribution
to sucrose metabolism
Figure 3
Vacuolar
invertases
Cell wall
invertases
Sucrose
synthases
Developmental progression
Vacuolar invertases:
Sink initiation and expansion
Maximal generation of vacuolar hexoses
- Expansion (linked to photosynthate availability)
- Respiration
- Hexose-based sugar sensing
(cell division and initial differentiation)
Cell wall invertases:
Continued sink initiation and expansion
Maximal generation of cell wall hexoses
- Reduction of turgor-based transport inhibition,
especially as volume increases to near maximum
- Turgor reduction in zones of phloem unloading
(localized action often in much of development)
- Respiration
- Hexose-based sugar sensing
(cell division and related gene expression)
Sucrose synthases:
Storage and maturation
Generation of UDPG instead of glucose
- Direct shuttling of UDPG to biosynthesis
- ATP-conserving respiratory path
- Minimization of hexose-based sugar sensing
(enhanced expression of genes for
storage and maturation)
Current Opinion in Plant Biology
A common developmental profile of the contributions of sucrosecleaving enzymes to sequential stages of sink initiation, expansion, and
storage/maturation. Both vacuolar and cell wall invertases maximize
hexose production, which in turn enhances respiration and the
generation of hexose-based signals. These hexose-based signals
upregulate diverse early development genes, including those
involved in cell division. Vacuolar invertases concurrently favor cell
expansion through their two-for-one enhancement of osmotic
solutes in the vacuoles, as well as by linking the initial volume
increase to photosynthate availability. As cell expansion slows, the role
of cell wall invertase becomes increasingly important in preventing the
turgor-based inhibition of symplastic transfer, and also in reducing
turgor in the zones of phloem unloading. As the cell division and
expansion phases shift to those of storage and maturation, sucrose
synthases become more important, and contribute in at least three
overall areas. These are their capacity to direct carbon to an ATPconserving respiratory path that is advantageous in many sink
tissues, to facilitate the shuttling of UDPG to biosynthetic products
or other special functions, and to minimize the production of hexoses
and thus hexose-based signals during specific stages of sink
development.
www.sciencedirect.com
Figure 4
(a)
Sucrose synthase
Sieve-tube element
Open
lumen
Companion cell
(b)
Cell
wall
Sieve-tube cytoplasm
Sievetube
lumen
Sucrose
UDP
Sucrose synthase
Biosynthesis
Respiration
UDPG Fructose
PPi
Callose
synthase
UGPase
PPi
F6P
UTP
G1P
Glycolysis
?
Callose
for plugging
ATP
for transporter
ATP
ADP
Sucrose
H+
H+
Figure 5
Plasma
membrane
Cytoplasm
Cellulo
se
Sucrose
Sucrose synthase
Cellulose synthase complex
Plasma membrane
ose
llul
Ce
Pore
subunit
Cytoplasm
UDPG
UDPG
UDP
UDP
Cellulose
synthase
Fructose
Sucrose
Amyloplasts
Actin
Sucrose
synthase
Cell wall
biosynthesis
(Special functions)
(c)
Cytoplasmic
Membrane
-bound
Plasma membrane
Golgi
SUS
(Respiration)
Tonoplast
Vacuole
Despite the extreme stability of sucrose synthase protein under some conditions [77], it is becoming apparent
that several mechanisms contribute to tightly control its
turnover (Figure 6). First, the phosphorylation that activates the enzyme (at the S15 site or its equivalent) [86,87]
is also implicated in predisposing sucrose synthase to
phosphorylation at a second site (S170 or its equivalent)
[90]. This second phosphorylation, in turn, targets the
protein for ubiquitin-mediated degradation via the proteosome [90]. Phosphorylation at the first site can be
linked to sugar availability and/or to other signals through
the catalysis of this step by both Ca2-dependant protein
kinases (CDPKs) [87,90,91] and/or a sugar-responsive
Ca2-independent SnRK [75,88,89,90]. The second
phosphorylation can be catalyzed by CDPKs but not
SnRKs [90]. Second, this avenue of sucrose synthase
breakdown can be inhibited if the second phosphorylation site (S170 or equivalent) is blocked by the binding of
ENOD40 proteins (which were initially named for their
role in early nodule development but which are now
known to have diverse functions throughout the plant)
[90,92,93]. The protective association of ENOD40s
with sucrose synthase has been implicated in the control
of vascular function, phloem loading/unloading, and
www.sciencedirect.com
Figure 6
Stable SUS
S15 S170
SUS
Activated
by +P
S15 S170
S15
ENOD
proteins
S170
SUS
SUS
Unstable SUS
P
P
S15
S170
+ P by CDPKs
SUS
S15 S170
Ubiquination and
degradation by proteosomes
SUS
It is now becoming evident that vacuolar invertases contribute prominently to both sucrose import ([5,17,36];
P Commuri et al., unpublished data) and sugar signals
[9,27,32], particularly during the expansion growth of
diverse sink structures [1,5,9,17,26,36,95]. These roles
for vacuolar invertases are additional to previously recognized functions including turgor regulation and the control of sugar balance in fruit tissues and mature tubers
[5,9,96]. The sucrose import and potential signaling
influences of vacuolar invertases arise from their role in
www.sciencedirect.com
Figure 7
Vacuole
Ribosomes
PPV
VIN
active?
VPE
inactive
VIN
VIN transfer and extended
transcription
protective storage
VIN
active
VIN
degraded
VPE
auto-activated
Regulation of vacuolar invertase (VIN) at the protein level. The transfer, protective retention, and protein turnover of VIN are potentially modulated by
the precursor protease vesicle (PPV) and vacuolar processing enzyme (VPEg) system. At least some of the newly translated VIN enters the PPV,
where it can be retained for extended periods. It is presumably protected from proteolytic degradation in this compartment, but its functional
role remains unclear until it enters the vacuole. The PPV also stores a VPEg protease, which remains inactive until it too moves into the vacuole.
Once activated, VPEg targets specific substrates that include a VIN. This invertase may thus be targeted for degradation as soon as it enters the
site of its presumed action in vivo. The PPV and VPEg system therefore provides a potential mechanism for modifying both the timing and the
duration of at least some VIN activity.
Current Opinion in Plant Biology 2004, 7:235246
www.sciencedirect.com
that a maize invertase inhibitor, ZmINVINH1, is exported to the apoplast, where it could interact with invertases
during early kernel development (i.e. 47 days after
pollination). These researchers further defined the most
prominent site of ZmINVINH1 expression as the embryo
surrounding region (ESR), which they suggest may help
to preserve crucial differences in the extracellular sugar
environments of the embryo and endosperm during early
development. Greater hexose levels in the endosperm
apoplast may favor more rapid cell division and minimal
differentiation relative to those in embryos [6].
Conclusions
Sucrose-cleaving enzymes lie at the heart of mechanisms
for the distribution and use of sucrose within multicellular
plants. They also occupy a pivotal position in the balance
between the different sugar signals generated by
imported sucrose. Their regulation has thus become
the focus of considerable interest, and involves diverse
and highly integrated mechanisms operating at transcriptional and posttranscriptional levels.
Acknowledgements
Supported by the US National Science Foundation (Cellular
Biochemistry) and by the Florida Agricultural Experiment Station
(Journal Series Number R-10079).
2.
3.
4.
5.
6.
7.
8.
Koch KE, Ying Z, Wu Y, Avigne WT: Multiple paths of sugarsensing and a sugar/oxygen overlap for genes of sucrose and
ethanol metabolism. J Exp Bot 2000, 51:417-427.
9.
38. Gardiner JC, Taylor NG, Turner SR: Control of cellulose synthase
complex localization in developing xylem. Plant Cell 2003,
15:1740-1748.
39. Salnikov VV, Grimson MJ, Seagull RW, Haigler CH: Localization of
sucrose synthase and callose in freeze-substituted secondarywall-stage cotton fibers. Protoplasma 2003, 221:175-184.
40. Ruan YL, Llewellyn DJ, Furbank RT: Suppression of sucrose
synthase gene expression represses cotton fiber cell initiation,
elongation and seed development. Plant Cell 2003, 15:952-964.
An interesting gradation of phenotypes indicates that antisense reductions
of sucrose synthase at the seed surface inhibit the development of
trichomes, and that internal reductions have still more pronounced effects.
The data support an association between sucrose synthase and cell-wall
biosynthesis, as well as other developmental roles for sucrose synthase.
41. Yanaglsawa S, Yoo S-D, Sheen J: Differential regulation of EIN3
stability by glucose in ethylene signaling in plants. Nature 2003,
425:521-525.
42. Gibson SI: Sugar and phytohormone response pathways:
navigating a signaling network. J Exp Bot 2004, 55:253-264.
43. Leon P, Sheen J: Sugar and hormone connections. Trends Plant
Sci 2003, 8:110-116.
28. Moore B, Zhou L, Rolland F, Hall Q, Cheng WH, Liu YX, Hwang I,
Jones T, Sheen J: Role of the Arabidopsis glucose sensor HXK1
in nutrient, light, and hormonal signaling. Science 2003,
300:332-336.
44. Lu C-A, Ho T-HD, Ho S-L, Yu S-M: Three novel MYB proteins with
one DNA binding repeat mediate sugar and hormone regulation
of a-amylase gene expression. Plant Cell 2002, 13:1963-1980.
Direct probe analyzes indicate that low oxygen is common in the phloem
environment and has direct implications for phloem function.
56. Ricard B, Toai VT, Chourey P, Saglio P: Evidence for the critical
role of sucrose synthase for anoxic tolerance of maize roots
using a double mutant. Plant Physiol 1998, 116:1323-1331.
57. Fukao T, Kennedy RA, Yamasue Y, Rumpho ME: Genetic and
biochemical analysis of anaerobically induced enzymes during
seed germination of Echinochloa crus-galli varieties tolerant
and intolerant of anoxia. J Exp Bot 2003, 54:1421-1429.
58. Gordon AJ, Minchin FR, James CL, Komina O: Sucrose synthase
in legume nodules is essential for nitrogen fixation.
Plant Physiol 1999, 120:867-878.
59. Xie ZP, Staehelin C, Broughton WJ, Wiemken A, Boller T, Muller J:
Accumulation of soluble carbohydrates, trehalase and sucrose
synthase in effective (Fix(R)) and ineffective (Fix(S)) nodules
of soybean cultivars that differentially nodulate with
Bradyrhizobium japonicum. Funct Plant Biol 2003, 30:965-971.
60. Blee KA, Anderson AJ: Transcripts for genes encoding soluble
acid invertase and sucrose synthase accumulate in root tip
and cortical cells containing mycorrhizal arbuscules. Plant Mol
Biol 2002, 50:197-211.
61. Hohnjec N, Perlick AM, Pu hler A, Kuster H: The Medicago
truncatula sucrose synthase gene MtSucS1 is activated both in
the infected region of root nodules and in the cortex of roots
colonized by arbuscular mycorrhizal fungi. Mol Plant Microbe
Interact 2003, 16:903-915.
62. Ravnskov S, Wu Y, Graham JH: Arbuscular mycorrhizal
fungi differentially affect expression of genes coding for
sucrose synthases in maize roots. New Phytol 2003,
157:539-545.
63. Pien S, Wyrzykowska J, Fleming AJ: Novel marker genes for early
leaf development indicate spatial regulation of carbohydrate
metabolism within the apical meristem. Plant J 2001,
25:663-674.
65. King SP, Lunn JE, Furbank RT: Carbohydrate content and
enzyme metabolism in developing canola siliques.
Plant Physiol 1997, 114:153-160.
83. Buckeridge MS, Vergara CE, Carpita NC: Insight into multi-site
mechanisms of glycosyl transfer in (1!4)beta-D-glycans
provided by the cereal mixed-linkage (1!3),(1!4)beta-Dglucan synthase. Phytochemistry 2001, 57:1045-1053.
www.sciencedirect.com
90. Hardin SC, Tang G-Q, Scholz A, Holtgraewe D, Winter H, Huber SC:
Phosphorylation of sucrose synthase at serine 170: occurrence
and possible role as a signal for proteolysis. Plant J 2003,
35:588-603.
The authors describe a previously unrecognized phosphorylation site of
sucrose synthase and its potential role in controlling the turnover of the
sucrose synthase protein.
91. Cheng SH, Willmann MR, Chen HC, Sheen J: Calcium signaling
through protein kinases. The Arabidopsis calcium-dependent
protein kinase gene family. Plant Physiol 2002, 129:469-485.
92. Kouchi H, Takane K, So RB, Ladha JK, Reddy PM: Rice ENOD40:
isolation and expression analysis in rice and transgenic
soybean root nodules. Plant J 1999, 18:121-129.
93. Rohrig H, Schmidt J, Miklashevichs E, Schell J, John M: Soybean
ENOD40 encodes two peptides that bind to sucrose synthase.
Proc Natl Acad Sci USA 2002, 99:1915-1920.
A combination of in-vitro translation and western blotting indicates that
two small ENOD40 peptides are produced from overlapping regions at
the 50 end of the same mRNA. Further work demonstrates that these
peptides bind specifically to sucrose synthase. This work, together with
data from other studies [90,94], implicates these ENOD40 peptides in
enhancing the activity and longevity of sucrose synthase in diverse
systems.
94. Varkonyi-Gasic E, White DW: The white clover enod40 gene
family. Expression patterns of two types of genes indicate a
role in vascular function. Plant Physiol 2002, 129:1107-1118.
www.sciencedirect.com