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Chapter 3 : System of Subsystems

3.6 Summary: A System with Subsystems

Guard cell signalling is a complex system that is coherently organized to control the stomatal aperture. These are signal perception, sphingolipid signalling, inositol signalling, ion

channels, ion channel regulatory proteins, Ca2+ signalling and NO signalling. This system

comprises an interconnected set of subsystems made of proteins, lipids, small molecules and various other conditions of the guard cells. The interconnections among the subsystems facilitate the communication flow of the whole system to respond to the signals by means of a collection of feedback processes and their relative dominance in time and space.

There are seven self-organized functional subsystems linked together to carry the ABA signal to close stomata. In systems thinking, this tight binding by feedback loops, which function differently in time and space with a defined hierarchy, provides the ABA network with better resilience to function in a variable environment with minimum time delay.

Ion channel signalling and Ca2+ signalling, being hubs in the system, are tightly connected

with many of the other subsystems. Of these two, the largest number of positive and/or negative

feedback loops connect Ca2+ signalling to the majority of subsystems, placing it as the core

reinforcement in the system.

Table 3.1: Regulatory mechanisms of ABA signalling network considered in the wiring diagram in Figure 3.5 (coloured according to modules in Figure 3.5)

Node

Regulation Target Source

Sphingolipid Signalling

SK ABA Not clear

SK PA Increase the specificity constant to promote substrate binding S1P SK Phosphorylation of Sphingosine to produce S1P

GPA1 S1P Addition of GTP permitting PLD release

PLD GPA1 Binding of GTP to GPA1 (Gα-GTP)dissociates GPA1 from PLD PLD NO May be S-nitrosylation of Cys residues or nitration of Tyr residues PLD CA Strengthen enzyme-substrate binding/targeting membrane localization of proteins

PA PLD Facilitates the hydrolysis of P-choline to produce PA

RCN1 PA Not clear

Signal Perception via ABA

PYR ABA Physical binding (conformational changes)

CuAO ABA Induce biosynthesis

PI4P ABA Phosphoinositide turnover PI4P PA Phosphoinositide turnover PI3P ABA Phosphoinositide turnover

PIP2 PI4P Phosphorylation

Inositol Signalling PLC ABA Possibly via phosphorylation

PLC CA Ca2+ driven activity and membrane targeting

InSP3 PLC

PLC hydrolyse PIP2 to produce InSP3

InSP3 PIP2

InSP6 InSP3 Phosphorylation by respective kinases InSP6 MRP5 Transport InSP6 to the vacuole

Osmoregulatory Enzymes/molecules

pH SnRK2 Not clear

PP2C PYR Physical binding (conformational changes)

PP2C PA Decreases the phosphatase activity and/or membrane-tethering role

PP2C ROS Oxidization

PP2C GPX3 Possibly by oxidation

PP2C pH Regulating phosphatase activity

PP2C ROP11 Physically binds to protect from Receptor inhibition

PP2C ABH1 Not clear

ROP11 ABA Not clear

ROP11 ERA1 Changing localization

SnRK2 PP2C Dephosphorylation

MAPK ROS May be through post-translational modification of redox sensitive proteins (cysteine oxidation)

MAPK CDPK Not Known

MAPK CA Not Known

MAPK PP2C Dephosphorylation

CDPK CA Ca

2+ binding to the N-terminal lobe

separates auto inhibitory domain from the active site

CDPK PP2C Dephosphorylation

RbOH SnRK2 Phosphorylation

RbOH CDPK Phosphorylation

RbOH CA Cabinding to ROS production is not clear 2+ binds to EF-hand motif of RbOH but the significance of this

RbOH RCN1 Not clear

RbOH PA Physically binds and stimulates

RbOH PI4P Not clear

RbOH PI3P Not clear

RbOH pH Not clear

ROS CuAO Catabolism of CuAO (oxidation) ROS RBOH O2 RBOH �⎯⎯⎯⎯⎯⎯⎯�O2-(ROS)

ROS GPX3 H2 O2 GPX3 �⎯⎯⎯⎯⎯⎯⎯⎯� H2O

GPX3 ROS High levels of system ROS lead the activation of the ROS scavenging NO Signalling

NO ROS Not clear

NO CaM Ca2+- bound CaM interact NOS like Enzyme to produce NO

CADPR CGMP

β-NAD+ ADP-ribosyl cyclase�⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯�cADPR

Activate ADP-ribosyl cyclase via G-kinase to stimulate cADPR synthesis

CGMP NO Activates soluble guanylyl cyclase resulting cGMP by nitrosylation

Ion Channels SLAC1 SnRK2 Phosphorylation

SLAC1 CDPK Phosphorylation

SLAC1 PP2C Dephosphorylation

SLAC1 MAPK Not clear

SLAC1 ERA1 Protein farnesylation

SLAC1 MRP5 Not clear

SLAC1 Malate Increasing current noise

TPC1 CA Ca2+ induced Ca2+ release by binding to EF-hands TPC1 DEPOLAR Voltage gated regulation

GORK NO Nytrosylation

GORK pH Membrane delimited pathway

GORK ROS Post-translational modification increasing current intensity GORK DEPOLAR Setting active potential for channel opening

QUAC SnRK2 Phosphorylation

QUAC DEPOLAR Voltage gated regulation

ATALMT6 CA Ca2+-dependent current activation

KAT1 CDPK Phosphorylation

KAT1 SnRK2 Phosphorylation

QUAC Malate Malate is a substrate for R-type ion channels

TPK1 CA Conformational changes induced by binding into EF-hands

TPK1 pH Not clear

Malate PEPC

Catalyses the b-carboxylation of PEP to yield oxaloacetate (OAA) and inorganic phosphate (the branch-point step in the malate- accumulation pathway)

PEP(C3) → OAA(C4) → Malate

Malate QUAC Transport malate Malate ATALMT6 Transport malate

AHA1 PP1 Through phosphorylation, displace C-terminal auto inhibitory domain AHA1 CA Post-translational modulation

AHA1 pH Not clear

Calcium Signalling CA Icas Influx of Ca2+ to cytosol CA CADPR Influx of Ca2+ to cytosol CA Ica Influx of Ca2+ to cytosol CA InSP6 Influx of Ca2+ to cytosol

CA Ca2+-ATPase Efflux of Ca2+ from cytosol

CA CAX1 Efflux of Ca2+ from cytosol

CaATPase CaM Suppress auto-inhibitory action by binding into N-terminus CaM CA Ca2+ binding enhances catalytic activity

Ica ABH1 Not known

Ica ERA1 May be through farnesylation

Ica MRP5 Not Known

Ica ROS ROS modify Ica channel proteins through directly and/or through additional intermediate proteins Ica DEPOLAR Changes in electrical potential

CBL CA Ca2+ binding for activation CAX1 CIPK May mask auto-inhibitory domain

SCAB1 ABA Not clear

SCAB1 InSP6 Not clear

Icas ACTIN Stretch activation

CIPK CBL Mask auto-inhibitory domain and facilitate localization CIPK PP2C Replace CBL protein to enhance auto inhibition PP1 PA Inhibits the phosphatase activity

ABPS CA Capping and depolymerization

ABPS PI3P Inactivate actin stabilization and facilitate depolymerization

ABPS PI4P

ACTIN ARP23 Actin nucleation

ACTIN SCAB1 Stabilizes actin filaments

ACTIN ATRAC1 Not clear

ACTIN ROS Depolymerize actin filaments via bonds weakening of inter-monomer ACTIN CA Facilitate depolymerization

ACTIN ABPS Stabilize actin filaments

PEPC Malate Binding to inhibitory site (feedback inhibitor-binding site)

ATRAC1 PP2C Not clear

ARP23 PIP2 PIP2 concentration may induce a ‘switch’ for N-WASP mediated Arp2/3 actin polymerization

ARP23 ABA Not clear

DEPOLAR CA Accumulation of positive ions makes membrane more positive DEPOLAR TPK1 Addition of K+ to cytosol from the vacuole

DEPOLAR AHA1 Facilitate membrane hyperpolarization

DEPOLAR SLAC1 Removal of negative charges makes membrane more positive

DEPOLAR QUAC Removal of negative charges makes membrane more positive CLOSURE GORK Removal of K+ (osmotic regulation)

CLOSURE ACTIN Rearrangement of cytoskeleton

CLOSURE Malate Removal of malate (osmotic regulation) CLOSURE SLAC1 Removal of Cl- (osmotic regulation)

Chapter 4

: Mathematical Framework: Boolean