Cell Signalling in Plants

Ms. Divya Mistry

Introduction
• Each kingdom has independently invented signalling mechanisms to
regulate growth and to pattern tissues.
• Plant signalling involves the conveying of information within and
between plant cells from receptor systems to effectors.
• Signalling can occur locally within a single plant or between different
plants, including plants of different species.
• The initial response of an organism to a signal takes place at the level
of the single cell.
What is Signalling ?
• Cell signalling is an essential, ubiquitous process that living systems use to
respond to the environment.
• Cell signalling includes cellular decisions such as development, cell growth and
division, differentiation, migration, apoptosis etc.
• Understanding cell signalling is critical, due to its importance in cellular fate
decisions and because malfunctions in cellular signalling are at the heart of many
diseases.
• Communication between cells is mediated mainly by extracellular signal
molecules.
• Signalling operates over long distances or to immediate neighbours/ short
distances.
• Multicellular organisms both emit and receive signals.
• Reception of the signals depends on receptor proteins at the cell surface, which
bind the signal molecule. The binding activates the receptor, which in turn
activates one or more intracellular signalling pathways.
• There is relay chains of which process the signal inside the receiving cell and
distribute it to the appropriate intracellular targets.
• The targets are generally effector proteins, which are altered when the signalling
pathway is activated and implement the appropriate change of cell behaviour.
• Depending on the signal and the nature and state of the receiving cell, these
effectors can be gene regulatory proteins, ion channels, components of a metabolic
pathway, or parts of the cytoskeleton.
What Kind of Signals Do Cells Receive?
• Most cell signals are chemical in nature.
• In multicellular organisms, growth factors, hormones, neurotransmitters,
and extracellular matrix components are some of the types of chemical
signals cells use. E.g. neurotransmitters are a class of short-range signalling
molecules that travel across the tiny spaces between adjacent neurons or
between neurons and muscle cells.
• Other signalling molecules move much farther to reach their targets. E.g.
follicle-stimulating hormone, which travels from the mammalian brain to
the ovary, where it triggers egg release.
• Some cells also respond to mechanical stimuli. E.g. sensory cells in the skin
respond to the pressure of touch, cells in the ear react to the movement of
sound waves etc.
How do Cells recognize signals?
• Cells have proteins called receptors that bind to signalling molecules and initiate a
physiological response.
• Dopamine receptors bind dopamine, insulin receptors bind insulin, nerve growth
factor receptors bind nerve growth factor, etc.
• There are hundreds of receptor types found in cells, and varying cell types have
different populations of receptors.
• Receptors can also respond directly to light or pressure, which makes cells
sensitive to events in the atmosphere.
• We know a group of similar cells working together is called a tissue.
How does that happen?
• When a molecule within the matrix
binds to the receptor, it changes the
molecular structure of the receptor.
• The receptor, in turn, changes the
conformation of the microfilaments
positioned just inside the plasma
membrane.
• These conformational changes induce
chemical signals inside the cell that
reach the nucleus and turn “on” or “off”
the transcription of specific sections of
DNA, which affects the production of
associated proteins, thus changing the
activities within the cell.
Intercellular Junctions
• Cells can also communicate with each other via direct contact, referred
to as intercellular junctions.
• Plasmodesmata are junctions between plant cells, whereas animal cell
contacts include tight junctions, gap junctions, and desmosomes.
Plasmodesmata
• Plasmodesmata (singular = plasmodesma),
numerous channels that pass between cell
walls of adjacent plant cells, connect their
cytoplasm, and enable materials to be
transported from cell to cell, and thus
throughout the plant.
• A gap in the cell wall, a plasmodesma,
allows fluid and small molecules to pass
from the cytoplasm of one cell to the
cytoplasm of the other.
 Tight junctions, Gap junctions, and Desmosomes

• Tight junctions: A tight junction is a watertight seal between two adjacent animal cells. The cells are held
tightly against each other by proteins called claudins and occludins.
• Gap junctions: Gap junctions in animal cells are like plasmodesmata in plant cells in that they are channels
between adjacent cells that allow for the transport of ions, nutrients, and other substances that enable cells
to communicate.
• Desmosomes : These act like spot welds between adjacent epithelial cells. Short proteins called cadherins
in the plasma membrane connect to intermediate filaments to create desmosomes.
What is a Ligand?
• Ligands are small molecules that transmit signals in between or within cells.
• Ligands exert their effects by binding to cellular proteins called receptors.
• A ligand is a protein that attaches (binds) to another protein called a receptor;
receptor proteins have specific sites into which the ligands fit like keys into locks.
• There are two main types of ligands: ligands that bind to receptors inside the cell,
called intracellular ligands, and ligands that bind to receptors outside the cell,
called extracellular ligands.

(Endogenous ligands are those that are produced in the body, not exogenous like
those introduced into the body, such as certain drugs.)
What a drug does to the body?
• A drug is any exogenous chemical / a ligand which affects the workings of body.
• Together, ligands and their receptors trigger signals that affect cell development
and function.
• Alterations in ligands can impair cell signalling and change the normal activities
of cells.
• Most drugs “act” on receptors (there are some that don’t). Receptors are molecules
(usually proteins) to which specific substances can bind and cause effect.
• A drug thus becomes an exogenous ligand, i.e. it comes from outside the body.
However, many molecules in the body work by binding to receptors too; these
are therefore called endogenous ligands, i.e. they come from inside the body.
When we think about how they cause their
effect...... drugs can be referred to as agonists or
antagonists.
• Drugs targeted to membrane receptors can have a variety of effects.
• They may elicit/stimulate or produce the same biological effects as the
natural ligand. If so, they are called agonists.
• An agonist is a mimetic of the natural ligand and produces a similar biological effect
as the natural ligand when it binds to the receptor.
• When receptors bind their natural target ligands (hormones,
neurotransmitters), a biological effect is elicited.) Conversely they may
inhibit/ obstruct or stop the biological activity of the receptor. If so they
called antagonists.
An antagonist inhibit the effects of the natural ligand (hormone, neurotransmitter).
• Agonists can be subdivided further to full, partial, and inverse agonists.
Agonists can be subdivided
further to full, partial, and
inverse agonists.
• A full agonist will produce a full response,
i.e. 100% of the response seen by
endogenous ligands. Using our analogy,
the lock is fully opened.
• Partial agonists stimulate the receptor to a
limited extent, but in doing so prevent any
further stimulation from endogenous
substances.
• An inverse agonist is one which reverses
constitutive activity of a receptor. An
inverse agonist does exert an effect itself,
and so is an agonist.
Antagonists can also be
subdivided into competitive or
non-competitive, reversible or
irreversible.
• A competitive antagonist competes with
the substrate to bind to the receptor.
• A non-competitive antagonist inhibits the
response by binding to an area separate to
the active site of the receptor.
• Reversible and irreversible inhibition is
pretty easy to explain. Either it can be
reversed…or it cannot.
What are Receptors?
• Receptors are special structures that can be found in cell membranes.
• These are made of protein molecules such as glycoproteins.
• Receptors are generally transmembrane proteins, which bind to signalling molecules
outside the cell and subsequently transmit the signal through a sequence of molecular
switches to internal signalling pathways.
• They receive signals and initiate a response, by binding to signalling molecules, often
called first messengers or ligands.
• A specific ligand will have a specific receptor that typically binds only that ligand.
• Cell surface receptors (membrane receptors, transmembrane receptors) take part in
communication between the cell and the outside world.
• Extracellular signalling molecules (usually hormones, neurotransmitters, cytokines,
growth factors or cell recognition molecules) attach to the receptor.
Types of Signalling
What is the Need?
A cell within a multicellular organism need to
signal to other cells that are at various
distances from the original cell to parse the
message.
There are four categories of chemical
signalling found in multicellular organisms:
• autocrine signalling (targets itself )
• direct signalling (a cell connected by gap
junctions, involves signalling molecules
moving directly between adjacent cells)
• paracrine signalling (acts on nearby cell)
• endocrine signalling (targets distant cell )
Types of Receptors
• INTERNAL RECEPTORS

• CELL-SURFACE RECEPTORS
a. ION CHANNEL-LINKED
RECEPTORS
b. G-PROTEIN-COUPLED
RECEPTORS (G proteins, also
known as Guanine nucleotide-
binding proteins)
c. ENZYME-LINKED RECEPTORS
INTERNAL RECEPTORS
• Internal receptors, also known as intracellular or
cytoplasmic receptors.
• These are found in the cytoplasm of the cell and respond to
hydrophobic ligand molecules that are able to travel across
the plasma membrane.
• When the ligand binds to the internal receptor -
• A change in shape is triggered that exposes a DNA-binding site
on the receptor protein
• The ligand-receptor complex moves into the nucleus, then
binds to specific regions of the DNA and promotes the
production of mRNA from specific genes
• Internal receptors can directly influence gene expression.
They often control functions such as gene transcription.
CELL-SURFACE RECEPTORS
• Cell-surface receptors, also known as transmembrane receptors.
• These are proteins that are found attached to the cell membrane.
• These receptors bind to external ligand molecules.
• This type of receptor spans or lengthens or distances the plasma membrane and
performs signal transduction, in which an extracellular signal is converted into an
intercellular signal.
• Cell-surface receptors are also called cell-specific proteins or markers because
they are specific to individual cell types.
• Each cell-surface receptor has three main components:
• an external ligand-binding domain
• a hydrophobic membrane-spanning region
• an intracellular domain inside the cell
ION CHANNEL-LINKED RECEPTORS
• Ion channel-linked receptors bind a ligand and open a channel through
the membrane that allows specific ions to pass through.
• It has an extensive membrane-spanning region.
• When a ligand binds to the extracellular region of the channel, there is
a conformational change in the proteins structure that allows ions such
as sodium, calcium, magnesium, and hydrogen to pass through.
G-PROTEIN-COUPLED RECEPTORS
• G-protein-coupled receptors bind a ligand and activate a membrane protein called a G-
protein (i.e. Guanine nucleotide-binding proteins).
• The activated G-protein then interacts with either an ion channel or an enzyme in the
membrane.
• Once the G-protein binds to the receptor, the G-protein changes shape, becomes active,
and splits into two different subunits.
• One or both of these subunits may be able to activate other proteins as a result of change.
• G protein-coupled receptors (GPCRs) are a large family of cell surface receptors that
share a common structure and method of signalling.
• A GPCR is made up of a long protein that has three basic regions: an extracellular portion
(the N-terminus), an intracellular portion (the C-terminus), and a middle segment
containing seven transmembrane domains.
• GPCRs are diverse and bind many different types of ligands. One particularly
interesting class of GPCRs is the odorant (scent) receptors. There are about 800 of
them in humans, and each binds its own “scent molecule” – such as a particular
chemical in perfume, or a certain compound released by rotting fish – and causes a
signal to be sent to the brain, making us smell a smell.
• GPCRs are found in the cell membranes of a wide range of organisms, including
mammals, plants, microorganisms, and invertebrates.
• The existence of GPCRs was demonstrated in the 1970s by American physician
and molecular biologist Robert J. Lefkowitz. He shared it with Brian K. Kobilka.
ENZYME-LINKED RECEPTORS
• Enzyme-linked receptors are cell-surface receptors with intracellular domains that
are associated with an enzyme.
• In some cases, the intracellular domain of the receptor itself is an enzyme.
• When a ligand binds to the extracellular domain, a signal is transferred through the
membrane, activating the enzyme. Activation of the enzyme sets off a chain of
events within the cell that eventually leads to a response.
• Receptor tyrosine kinases (RTKs) are
a class of enzyme-linked receptors
found in humans and many other
species.
Mechanism, targets
and responses of all
three cell surface
receptors
How Viruses Recognize a Host?
• Viruses often bind to cell-surface receptors on the host cell.
For example, the virus that causes human influenza (flu) binds specifically to receptors on
membranes of cells of the respiratory system. Chemical differences in the cell-surface receptors
among hosts mean that a virus that infects a specific species (for example, humans) cannot infect
another species (for example, chickens).
• Viral reproduction invariably produces errors that can lead to changes in newly
produced viruses.
• Changes happen randomly and quite often in the reproductive cycle
of a virus, but the changes only matter if a virus with new binding
properties comes into contact with a suitable host.
• Scientists watch newly appearing viruses (called emerging viruses)
closely in the hope that scientific monitoring can reduce the
likelihood of global viral epidemics.
SECONDARY MESSENGERS
• Secondary messengers are molecules that relay signals
received by receptors on the cell surface to target
molecules in the cytosol or nucleus.
• They serve to amplify the strength of the signal.
• Binding of a ligand to a single receptor at the cell surface
may end up causing massive changes in the biochemical
activities within the cell.
• Many pathways involve second messengers, small, non-
protein molecules that pass along a signal initiated by the
binding of a ligand (the “first messenger”) to its receptor.
There are 3 major classes of secondary
messengers:
1. Cyclic nucleotides (e.g., cAMP- cyclic adenosine monophosphate
and cGMP- cyclic guanosine monophosphate- it is a cyclic
nucleotide derived from guanosine triphosphate (GTP). cGMP acts
as a second messenger much like cyclic AMP.)
2. Inositol phosphates i.e. inositol trisphosphate (IP3) and
diacylglycerol (DAG)
3. Calcium ions (Ca2+)
Cyclic nucleotides (e.g., Cyclic AMP (cAMP))
• This is second messenger used in many different cell
types, it is a small molecule made from ATP. In response
to signals, an enzyme called adenylyl cyclase converts
ATP into cAMP, removing two phosphates and linking the
remaining phosphate to the sugar in a ring shape.
• cAMP can activate an enzyme called protein kinase A
(PKA) --- phosphorylation --- pass along the signal.
• Protein kinase A is found in a variety of types of cells, and
it has different target proteins in each.
• This allows the same cAMP second messenger to produce
different responses.
 Inositol phosphates i.e. Inositol trisphosphate (IP3)
and Diacylglycerol (DAG)
• Phospholipids called phosphatidylinositols can be
phosphorylated, releasing two fragments that both act as
second messengers.
• One particularly important lipid in this group is PIP2.
• It is essential to the cell membrane of all animal and plant cells and acts
as a second messenger in a variety of signalling pathways.
• In response to a signal, an enzyme phospholipase C cleaves PIP2 into
two fragments, DAG and IP3, both act as second messengers.
• DAG stays in the plasma membrane and can activate a target
called Protein Kinase C (PKC), allowing it to phosphorylate its
own targets.
• IP3 diffuses into the cytoplasm and can bind to ligand-gated
calcium channels in the endoplasmic reticulum, releasing Ca2+
that continues the signalling cascade.
Calcium ions (Ca2+)
• For signalling purposes Ca2+ may be stored in
compartments such as the endoplasmic reticulum.
• In pathways that use calcium ions as a second
messenger, signalling events release a ligand that binds
to and opens ligand-gated calcium ion channels.
• These channels open and allow the higher levels of
Ca2+ that are present outside the cell (or in intracellular
storage compartments) to flow into the cytoplasm,
raising the concentration of Ca2+.
• Some proteins in the cell have binding sites for Ca2+
ions, and the released ions attach to these proteins and
change their shape (and thus, their activity)
Signal transduction pathway
• The change in the receptor sets off a series of signalling events.
• This series of reaction can eventually lead to a change in the cell's behaviour or
characteristics.
• This chains that relay intracellular signals is known as intracellular signal
transduction pathway.
• Signal transduction pathway is a set of chemical reactions in a cell that occurs
when a signalling molecule attaches to a receptor.
• Steps in the signal transduction pathway often involve the addition or removal of
phosphate groups which results in the activation of proteins.
• This creates a phosphorylation cascade, where one enzyme phosphorylates
another, which then phosphorylates another protein, causing a chain reaction.
3 Stages of Cell Signalling
• Reception: A cell detects and receives a
signalling molecule from outside the cell.
• Transduction: When the signalling molecule
binds the receptor it changes the receptor
protein in some way. This change initiates the
process of transduction.
• Response: Finally, the signal triggers a cellular
response.

Protein phosphatases are a group of proteins important to the

phosphorylation cascade. Protein phosphatases are enzymes
that can rapidly remove phosphate groups from proteins
(dephosphorylation) and thus inactivate protein kinases.
Plant Signalling
• Plant signalling involves the conveying of information within and between plant
cells from receptor systems to effectors.
• Signals can be chemical or electrical, and signalling can occur locally within a
single plant or between different plants.
• Plants have been shaped by evolution as immobile, autotrophic and multicellular
organisms.
• Plant growth is highly elastic, undetermined, and strongly influenced by external
conditions.
• In Plants, two component regulatory system and target of rapamycin (TOR)
signalling, links cell and organism growth with that of the environment and its
factors.
Two-Component Pathway
• Two-component signal transduction (TCST) systems are the principal means for
coordinating responses to environmental changes in bacteria as well as some
plants, fungi, protozoa, and archaea.
• Two-component systems typically consist of
• a membrane-bound histidine kinase (HK) (sensor kinase) that senses a specific
environmental stimulus, signalling cascade induces change in cellular
behaviour and,
• the corresponding response regulator (RR) (DNA-binding response regulator)
which facilitates the cellular response, (through differential expression of
target genes)
• It helps Plants respond to outside stimuli by initiating signalling cascades that
regulate gene expressions.
• Organisms like bacteria, plants, fungi,
protozoa, and archaea respond to changing
environments by altering gene expression.
• Two-component regulatory systems (TCRS)
- important mediators/ intermediates of
signal transduction - enable organisms to
detect physical / chemical changes and relay
this signal through the cytoplasm to the
nucleus, where modulation of gene
expression occurs.
TCRS
• In its most basic form, a TCRS consists of a membrane-bound sensor kinase and a
DNA-binding response regulator.
• In response to specific stimuli, the sensor kinase is phosphorylated at a conserved
histidine residue and then the phosphoryl group is transferred to the conserved
aspartate on the response regulator.
• Phosphorylation of the response regulator triggers a conformational change,
driving dimerization and high affinity DNA binding.
• In prokaryotes and plants, signal transduction mediates by two component
regulatory system (TCRS), which is involved in cell-cell communication and to
respond extracellular signal.
• Two component system contain a sensor,
which is a homodimeric transmembrane
protein called Histidine kinase it has
autophosphorylating activity along with
conserved histidine residue and a response
regulator which has a conserved Aspartate
residue.
• Histidine kinase (HK) has two domains one is
histidine phosphotransfer domain, which
possesses specific histidine and second ATP
binding domain.
• Response regulator also has two domains one
conserved receiver domain which comprises
conserved Aspartate and second effector
domain.
Histidine kinase is also present in hybrid form called hybrid histidine kinase, in which
histidine kinase also contain one internal receiver domain, as ligand binds to hybrid
histidine kinase it autophosphorylates itself and sets further cascade of events.

Schematic representation and basic features of the simple (A) and the multistep (B) two-component signaling
systems.
• Two-component systems have emerged as
important response mechanisms in higher
plants.
• Composed of hybrid histidine kinases,
histidine-containing phosphotransfer domain
proteins and response regulators that are
biochemically linked by His-to-Asp
phosphorelay mech.
• In plants two-component systems play a major
role in cytokinin perception, ethylene signal
transduction, osmosensing, developmental
processes – megagametogenesis (Arabidopsis
thaliana) -flowering promotion (Oryza sativa)
etc.
• Two-component-like elements also function as
components of circadian clock (Arabidopsis
thaliana)
Tor Signalling networks in Plants
• Target of Rapamycin (TOR) is an evolutionarily conserved protein kinase (TOR
kinase).
• It is a highly conserved serine/threonine kinase that controls cell growth and
metabolism in response to nutrients, growth factors, cellular energy, and stress.
• Rapamycin, also known as Sirolimus, is a macrolide compound that is used to coat
coronary stents, prevent organ transplant rejection and to treat a rare lung disease
called lymphangioleiomyomatosis.
• It is a type of antibiotic, a type of immunosuppressant, and a type of
serine/threonine kinase inhibitor.
Tor Signalling networks in Plants
• Rapamycin was initially discovered as an antifungal metabolite produced
by Streptomyces hygroscopicus from a soil sample of Easter Island.
• Rapamycin has immunosuppressive and anti-proliferative properties in
mammalian cells and is especially useful in preventing the rejection of kidney
transplants.
• It was isolated for the first time in 1972 by Surendra Nath Sehgal and colleagues.
• TOR takes part in nutrient, energy, hormone, growth factor cycles and
environmental inputs to control proliferation, growth and metabolism in diverse
multicellular organisms.
• It is said Mammalian target of rapamycin (mTOR) not only controls cell growth
and proliferation but also survival.
Rapamycin: The Antiaging Molecule
Studies
• After the discovery of rapamycin, studies showed its capacity to inhibit cancer cell
proliferation in mouse models, while some studies explored the potential of
rapamycin as an immunosuppressant for organ transplants.
• Rapamycin has also shown efficacy against mouse models of age-related diseases
(including cancer and neurodegenerative diseases including Alzheimer’s disease, age-related
cognitive decline).
• Rapamycin also showed to extend the lifespan of wild-type mice in 2009 by the
National Institute on Aging Interventions Testing Program (significant discussion and
research is needed whether it is a true antiaging intervention or “merely” a potent anticancer
agent).
TOR Kinase – central component
• Many studies indicate that TOR acts as a molecular switch for the activation of
cell proliferation or production and plant growth at the expense of cellular
immunity.
• It acts as a central component of TOR signaling and modifies several downstream
proteins by phosphorylation.
• Cell growth is an important process.
• Cells - sense environmental factors like nutrient abundance, the energy level or stress
signals and coordinate growth accordingly. The Target Of Rapamycin (TOR) pathway
is a major controller of these processes in all eukaryotes.
• Interfering with TOR signalling has a strong impact on plant development.
• The name of the TOR kinase and the entire pathway describes a characteristic
property – the specific and effective inhibition by rapamycin, a macrocyclic
lactone of bacterial origin.
Structure of the TOR protein
• The TOR protein belongs to the family of PIKK (phosphatidylinositol kinase-related
kinases) which represent a group of conserved serine/threonine kinases.
• In the N-terminal region, TOR consists of up to 20 tandem (trail like) HEAT repeats (The
name "HEAT" is an acronym for four proteins in which this repeat structure is found:
Huntingtin, elongation factor 3 [EF3], protein phosphatase 2A [PP2A], yeast kinase/ PI3-
kinase TOR1),
• FAT domain
• FRB domain (FKBP12-Rapamycin Binding domain of mTOR)
• the kinase domain and the FATC domain
TOR and Stress
• TARGET OF RAPAMYCIN (TOR) kinase controls
many cellular functions in eukaryotic cells in
response to stress and nutrient availability.
• Arabidopsis 3-phosphoinositide-dependent protein
kinase-1 (PDK1)
• Protein phosphatase 2A [PP2A]
• Ribosomal Protein S6 (RPS6) - S6 kinase (S6K)
• Gamma-butyrolactones (GBL)
This was also shown to be essential for embryonic development
in Arabidopsis thaliana. It was demonstrated that Arabidopsis
RAPTOR1 (a TOR regulatory protein) interacts with the HEAT
repeats of TOR and that RAPTOR1 regulates the activity of S6
kinase (S6K) in response to osmotic stress.
Plant TOR proteins
• TOR kinases from very diverse eukaryotic species show a high degree of
conservation in the kinase, FATC, and the FRB domain but only to a limited
extend in the number of HEAT repeats.
• The phylogenetic tree of TOR proteins from different species perfectly reflects
phylogenetic relationships of the species.
• Four main groups are visible corresponding to the animal kingdom, fungi, algae,
and higher plants.
• This strong conservation of TOR proteins throughout the species points out the
general importance of this kinase and, consequently, the entire TOR pathway.
Functional importance of the TOR kinase in
plants
• Both in plants and animals, TOR
exerts a very general function in
anabolic and catabolic processes.
• In Arabidopsis, TOR expression
is seen in early development in
the endosperm, the embryo and
the chalazal proliferating tissue.
Recollect

Ms. Divya Mistry