NPTEL Biotechnology -Systems Biology

Quorum Sensing - The Language of Bacteria

Dr. M. Vijayalakshmi
School of Chemical and Biotechnology
SASTRA University

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Table of Contents
1 INTRODUCTION .............................................................................................. 3
1.1 TYPES OF AUTOINDUCERS.............................................................................. 4
1.1.1 Acyl Homoserine Lactone molecules.................................................... 5
1.2 SYNTHESIS OF AUTOINDUCERS........................................................................ 6
1.3 PEPTIDE PHEROMONES- AUTOINDUCERS IN GRAM-POSITIVE BACTERIA ............... 7
1.4 BIOLUMINESCENCE AS A PHENOTYPE OF QUORUM SENSING- THE LUX SYSTEM.... 7
1.5 PHENOTYPES IN QUORUM SENSING SYSTEMS ................................................. 10
2 REFERENCES ................................................................................................ 11
2.1 LITERATURE REFERENCES ........................................................................... 11

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1 Introduction
This chapter deals with an interesting developmental and behavioural process in
bacteria, a sophisticated communication system that regulates the behaviour of a
bacterial population. Biochemical and genetic experiments in 1960s and 1970s
have provided compelling evidence of this organized social behavior in bacteria
and other micro organisms which can perceive and respond to the presence of
neighboring populations. Quorum sensing is a term employed to describe this
density dependent phenomenon in bacteria. Quorum sensing controls gene
expression in response to cell density as in Fig 1 and regulates physiological
functions predominantly in Gram-negative and Gram-positive bacteria.

Fig 1. General representation of quorum sensing in regulating gene expression based on cell density

The term quorum sensing first introduced in a review by Fuqua et al., (Journal of
bacteriology, 1994) traced the threshold level of individual cell required to initiate
a population response in concert.

The phenomenon of quorum sensing is

accomplished by the accumulation of small chemical signalling moieties which

accumulate extracellularly in the system and induce a population dependent
phenotype in the system. Scientific investigations in bacterial quorum sensing
began with the identification of density dependent bioluminescence in the marine
symbiotic bacterium Vibrio fischeri and another free living bacteria called Vibrio
harveyi reported in Nealson and Hastings, Journal of Bacteriology 1970,
Microbiol.Rev, 1979.
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Both the species produce and respond to a signalling molecule called Acylated
Homoserine Lactone (AHL) which is called an auto inducer that shoe
extracellular accumulation with cell growth. When the concentration of AHL
crosses the threshold level it initiates a signal transduction cascade with the
population density, leading to the production of luciferase. In other words the
entire circuit relies on the intracellular production and export of AHL, whose
extracellular concentration grows with population density. This signalling
molecule is sensed and reimported into the cells facilitating the whole population
to respond to a critical cell density.
Engbrecht and Silverman (Cell 1983, PNAS 1984) identified isolated and cloned
genes responsible for the density dependant gene regulation in Vibrio fischeri
and the genes encoding the luciferase enzyme complex. Their experiments
demonstrated the involvement of two regulatory proteins LuxI and LuxR where
LuxI is responsible for the synthesis of the auto inducer AHL and LuxR is a
transcriptional activator protein which binds to the auto inducer and enables
transcription of the structural operon LuxCDABE of luciferase.

1.1 Types of Autoinducers

Microbially derived signalling molecules act as auto inducers in bacterial quorum
sensing. The Gram-negative bacteria use fatty acid derivatives called
Homoserine Lactones HSLs whose synthesis is dependent on LuxI homolog or
LuxR homolog encoding a transcriptional activator protein responsible for
detection of the cognate HSL and the resulting gene expression which results in
phenotypic changes. More than 30 species of Gram-negative bacteria use HSL
derivatives for the control of the cell density and hence the quorum sensing
phenomenon.
The Gram-positive bacteria use amino acids and short peptide derivatives for
quorum sensing.

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1.1.1 Acyl Homoserine Lactone molecules

The AHL signal molecules from different bacteria are related in structure, but
differ in the nature of the acyl side chain moieties attached to them.The acyl
group can vary from 4 to 14 carbons depending on the auto inducer. It also
possesses a hydroxyl group, a carbonyl group, it is either fully saturated or
contains a single carbon-carbon double bond. A significant number of microbial
acyl HSLs have even number of carbons in their acyl side chains (Table 1) and
are synthesized by different bacterial genera. Many bacterial species can
produce more than one type of Acyl Homoserine Lactone and the type of acyl
HSL produced by a particular species can be strain dependent.

Table 1

Diversity exhibited by AHL molecules

Structure

Name
N-butanoyl-1-L-homoserine
lactone
(C4-HSL)
N-(3-hydroxybutanoyl)-L
homoserine lactone
(3-hydroxy-C4-HSL)
N-hexanoyl-L-homoserine
lactone
(C6-HSL)
N-(3-oxohexanoyl)-Lhomoserine lactone
(3-oxo-C6-HSL)
N-octanoyl-L-homoserine
lactone
(C8-HSL)
N-(3-oxooctanoyl)-Lhomoserine lactone
(3-oxo-C8-HSL)

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1.2 Synthesis of Autoinducers

The Homoserines found in bacteria are intermediates of the methionine-lysinethreonine bio synthetic pathway. S-adenosylmethionine (SAM) is one of the
intermediates of methionine/homocysteine pathway as in Fig 2.

Fig 2. Putative biosynthetic scheme for acyl-HSL synthesis by LuxI-type protein

SAM and the acyl-acyl carrier protein (acyl-ACP) act as substrates for LuxI type
of enzymes. This is an acyl-ACP generated using fatty acid biosynthesis. The
LuxI proteins couple a specific acyl-ACP to SAM through the formation of an
amide bond between the acyl side chain of the acyl-ACP and the amino group of
homo cysteine moiety of SAM. HSL is formed due to the lactonization of the
ligated

intermediate

in

the

reaction

along

with

the

release

of

methylthioadenosine. The acyl side chain moiety of the acyl- ACP determines the
specificity of interaction of a particular type of LuxI protein with its correct acylACP and this specificity regulates the LuxI protein to produce only one type of
the autoinducer AI.

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1.3
Peptide pheromones- autoinducers in Gram-positive
bacteria
Quorum sensing systems in Gram-positive bacteria use post transcriptionally
processed small peptides as signalling molecules. Such peptides are secreted by
ATP binding cassette transporters and interact with membrane bound sensor
kinases to initiate signal transduction across the membrane. Some of these
peptides are transported into the cell by oligopeptide permeases where they
interact with intracellular receptors Streptococcus and Enterococcus

produce

unmodified linear peptides few other species synthesize cyclic peptides. These
are synthesized as unprocessed translation products and are later processed
and secreted into the external environment. Gram-negative bacteria utilize LuxR
proteins as autoinducers while Gram-positive bacteria utilize two component
adaptor response proteins for detecting autoinducers. Secreted peptide signals
are detected by two component sensor kinases. Interaction with the peptide
induces a series of phosphorylation events that phosphorylate the cognate
response regulator protein activating it and allowing it to bind to the target DNA.
This binding alters the transcription of genes regulated by quorum sensing.

1.4 Bioluminescence as a phenotype of quorum sensing- The

Lux system
Vibrio

fischeri

and

Vibrio

harveyi

are

marine

bacteria

which

exhibit

bioluminescence as a phenotype in quorum sensing. Such bacteria do not

spread light and are non luminescent when they are free living in sea water at
lower densities. In laboratory experiments when cultures of Vibrio fischeri are
grown to high cell densities the culture shows bioluminescence with a bluish
green light at 493 nm. This behavior explains an interesting symbiotic
relationship of the bacterion with species such as the Japanese pinecone fish
and squid species such as Euprymna scolopes. Normally the squid house the
bacteria Vibrio fischeri in a special organ called the light organ. When the density
of the Vibrio fischeri population in the light organ increase to the order of 1010 and
1011 cells per ml the squid displays bioluminescence in the light organ. This
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phenotype caused by Vibrio fischeri is exploited by the squid to exhibit a

phenomenon called counter illumination. Interestingly at night the squid controls
the intensity of light it projects downwards and eliminates the visible shadow
created by the moon light. Hence camouflaging itself from its predators lying
below the surface. This explains the perfect symbiotic relationship where the
squid provide nutrient to the Vibrio fischeri and the bacterium Vibrio fischeri
secures the squid from predators through the luminescence it produces due to
quorum sensing.

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Video to be integrated - Principle of Quorum Sensing

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1.5 Phenotypes in quorum sensing systems

In microorganisms like Gram-positive and Gram-negative bacteria employ
quorum sensing and regulate phenotypes like virulence against host, antibiotics,
competence, bioluminescence, cell division, bio film formation and a variety of
others all based on the threshold population size and cross talk within their own
community. For example Bacillus subtilis uses quorum sensing for sporulation
and for the development of genetic competence. Streptococcus pneumonia uses
a signal peptide for establishing competence for transformation.
Table 2 below lists the different phenotypes exhibited by quorum sensing
systems in Gram-negative and Gram-positive bacteria.

Table 2
Genus

Quorum sensing Phenotypes

Phenotype

Aeromonas hydrophila
Aeromonas salmonicida
Agrobacterium tumefaciens
Bacillus
Chromobacterium violaceum
Escherichia coli
Lactococcus and other lactic
acid bacteria

Extracellular protease, biofilm formation

Myxococcus
Nitrosomonas europaea
Pseudomonas aeruginosa

Development

Rhodobacter sphaeroides
Streptococcus
Streptomyces
Vibrio (photo bacterium) fischeri
Xenorhabdus nematophilus

Community escape

Extracellular protease
Conjugation
Competence, development
Antibiotics, violacein, exoenzymes, cyanide
Cell division
Bacteriocin production

Emergence from lag phase

Multiple exoenzymes, Xcp, RhlR, biofilm formation, cyanide,
RpoS, lectin, pyocyanin, rhamnolipid
Competence, virulence
Antibiotic production
Bioluminescence
Virulence, bacterial lipase

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2 References
2.1 Literature References
1. Fuqua et al., Quorum sensing in bacteria: the LuxR-LuxI family of cell
density-responsive transcriptional regulators, Journal of bacteriology (1994),
176, 269275.
2. K H Nealson and J W Hastings, Bacterial bioluminescence: its control and
ecological significance, Microbiol Rev. (1979), 43, 496518
3. Joanne Engebrecht, Kenneth Nelson, Michael Silverman,

Bacterial

bioluminescence: Isolation and genetic analysis of functions from Vibrio

fischeri, Cell (1983), 32, 773781.
4. Charu Gera and S. Srivastava, Quorum- sensing: The phenomenon of
microbial communication, Current science (2006), 90, 666-676.

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