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Lecture 22: Microbial cell-to-cell communication

Today:

  • Do bacteria communicate with each other?
  • How do bacteria communicate with each other?

(wikipedia)

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And now, social behaviors in microbes!

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How do social behaviors arise?

DNA/genes

cells

genome

community/colony

organisms

ecosystem

What common principles lead to more complex biological organization?

One way to think about this is to look at how complex behaviors arise in general:

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“Traditional” Darwinian view of evolution

Cells growing on a nutrient source

A mutant arises in a gene that allows faster growth under these conditions

The mutant has a fitness advantage and now the new copy of the gene becomes much more frequent in the population

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How do you reconcile this view with “social” behaviors?

Worker Bees

(wikipedia)

Multicellularity

(wikipedia)

One approach: redefine fitness as “inclusive”!

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Inclusive fitness theory: a model for social evolution

Recipient

Actor

social action

offspring

offspring

+ or -

+ or -

r, how related are Actor/Recipient (probability they share the gene(s) for the social action)?

r1, how related are Actor and its offspring?

r2, how related are Actor and Recipient’s offspring?

m, the change in actor’s expected offspring as a result of the action

n, the change in recipient’s expected offspring as a result of the action

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Inclusive fitness theory: a model for social evolution

Recipient

Actor

social action

offspring

offspring

+ or -

+ or -

r

r1

r2

m

n

Effect on Recipient’s expected offspring number (n)

Gain (n > 0)

Loss (n < 0)

Effect on Actor’s expected offspring number (m)

Gain (m > 0)

Loss (m < 0)

Cooperation

Altruism

Selfishness

Spite

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Hamilton’s Rule

A gene G is associated with a social action.

The gene will spread if the total change in frequency brought about by the social action is positive, or if the inclusive fitness is positive!

 

 

 

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Hamilton’s Rule

Bourke, Principles of Social Evolution

Hamilton’s Rule asserts that a social phenotype that hurts the fitness of the actor can still arise evolutionarily if it suitably increases the fitness of a recipient that’s closely related!

What “social behaviors” can bacteria engage in??

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Quorum Sensing: Bacterial Communication

Vibrio fischeri

Ocean-dwelling V. fischeri only produce luminescence at high cell density.

How?

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Luminescence of the Hawaiian Bobtail Squid

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V. fischeri live within E. scolopes tissue

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sand

shallow water

shadow

Hawaiian Monk Seal

luminescent V. fischeri (1011 cells / mL!!)

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The squid goes to bed in the morning

V. fischeri

V. fischeri mostly purged by pump during sleep

  • Low cell density > no luminescence!?
  • As the day goes on cells grow and divide
  • By nighttime again, they reach a critical density at which the become luminescent!

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Luminescence turns on at night

V. fischeri

V. fischeri mostly purged by pump during sleep

  • Low cell density > no luminescence!?
  • As the day goes on cells grow and divide
  • By nighttime again, they reach a critical density at which the become luminescent!

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How do V. fischeri turn on luminescence only at high density?

“Autoinducer” molecule

Low cell density > low concentration of autoinducers

High cell density > high concentration of autoinducers

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How do V. fischeri turn on luminescence only at high density?

“Autoinducer” molecule

Low cell density > low concentration of autoinducers

High cell density > high concentration of autoinducers

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How do V. fischeri turn on luminescence only at high density?

luminescence genes

autoinducer

LuxI

LuxR

Transcription of luminescence genes!

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Quorum Sensing

“Autoinducer” or “Quorum Sensing” molecule

Low cell density > low concentration of autoinducers

High cell density > high concentration of autoinducers

Activate multicellular behaviors

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Quorum Sensing molecules are species-specific

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Two major QS paradigms

Gram negative

Gram positive

QS molecules (AHL) pass through membrane

QS molecules (polypeptides) activate membrane receptors

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What kind of behaviors are regulated by quorum sensing?

Behaviors that would not be useful when there is not a large group of bacteria!

Luminescence in V. fischeri

competence

Competence in B. subtilis

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What kind of behaviors are regulated by quorum sensing?

Behaviors that would not be useful when there is not a large group of bacteria!

Virulence in S. typhimurium

Mating/DNA transfer in A. tumefaciens

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What makes a phenotype “Quorum Sensing”?

Regulated by a species-specific extracellular molecule

Clear density-dependence

Increasing density

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Quorum sensing has remarkable synthetic biology applications!

First, a non-quorum sensing example, a synthetic oscillator circuit:

E. coli

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A synthetic oscillating E. coli

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The normal repressilator is very noisy, bad

Behaviors of different cells under different conditions:

How can we make this system more robust and reliable?

What if we coupled many cells together to cooperate? Would that reduce the noisiness?

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A synthetic quorum sensing oscillator

luxI

produces AHL QS molecule

AHL

aiiA

degrades AHL QS molecule

AHL

yemGFP

AHL

reporters on QS activity

All QS-regulated!

LuxR

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How does the system work?

  1. At low cell density, the fluidic channel sweeps away AHL too fast for QS to kick in
  2. Once cells reach a critical density, there is enough AHL to activate a burst of luxI transcription, producing more AHL, and activating both yemGFP and aiiA
  3. AiiA proteins degrade AHL, turning off luxI, yemGFP, and also aiiA itself
  4. With less AiiAs degrading AHL, AHL can accumulate again, leading to another cycle

Should result in oscillations in the production of YemGFP. Does it?

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GFP

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Oscillation period should depend on flow rate

If fluid flow is inhibiting quorum sensing by removing AHL from the system, oscillations should become slower as flow rate increases because it will then take longer for the cells to build up the critical AHL concentration

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Oscillation period increases with flow rate

fluid flow velocity

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QS oscillations should propagate in space

AHL

Because AHL 1) can diffuse through space, and 2) is coupled to its own production, if a group of cells reaches critical AHL concentration and starts a QS pulse, it should propagate through space.

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QS oscillations should propagate in space

AHL

Because AHL 1) can diffuse through space, and 2) is coupled to its own production, if a group of cells reaches critical AHL concentration and starts a QS pulse, it should propagate through space.

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A model predicts spatial QS waves

Within one cell

Across a spatially extended population

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What have we learned?

  • Inclusive fitness is a framework to account for the evolution of group behaviors��
  • Bacteria participate in social behaviors activated by cell-to-cell signaling molecules���
  • The molecular tools for this communication can be repurposed for engineered cell-to-cell-signaling systems