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The V. fischeri Autoinducer. N -( b -ketocaproyl)- L -homoserine lactone. LuxI. Quorum Sensing. LuxR. LuxR. target gene. Acyl-Homoserine Lactone Autoinducers. V. fischeri P.aeruginosa A. tumefaciens E. carotovora. Density dependent luminescence Virulence factors

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slide3

The V. fischeri Autoinducer

N-(b-ketocaproyl)-L-homoserine lactone

slide4

LuxI

Quorum Sensing

LuxR

LuxR

target gene

slide6

V. fischeri

P.aeruginosa

A. tumefaciens

E. carotovora

Density dependent luminescence

Virulence factors

Conjugal transfer of Ti plasmid

Antibiotic synthesis

LuxI-LuxR Quorum Sensing Systems

slide7

LuxP

LuxR

Quorum Sensing in V. harveyi

AI-2

LuxQ

H

D

H

D

HTH

LuxU

LuxO

H

D

LuxN

AI-1

LuxCDABE

slide9

Bacillus anthracis

Bacillus halodurans

Bacillus subtilis

Borrelia burgdorferi

Campylobacter jejuni

Clostridium acetobolyticum

Clostridium difficile

Clostridium perfringens

Deinococcus radiodurans

Escherichia coli

Enterococcus faecalis

Haemophilus influenzae

Helicobacter pylori

Klebsiella pneumoniae

Lactococcus lactis

Leuconostoc oenos

Listeria monocytogenes

Neisseria gonorrhoeae

Neisseria meningitidis

Pasteurella multocida

Porphyromonas gingivalis

Proteus mirabilis

Salmonella paratyphi

Salmonella typhi

Salmonella typhimurium

Shewanella putrefaciens

Staphylococcus aureus

Staphylococcus epidermidis

Streptococcus gordonii

Streptococcus mutans

Streptococcus pneumoniae

Streptococcus pyogenes

Vibrio anguillarum

Vibrio cholerae

Vibrio harveyi

Vibrio vulnificus

Yersinia pestis

Bacteria That Contain luxS

slide10

luxS

metK

pfs

slide12

MetK

Pfs

Pfs

Page544

slide13

SAM Utilization

Met

MetK

SAM

SAH

MTA

Pfs

Pfs

SRH

MTR

4,5-Dihydroxy-

2,3-pentanedione

+ HC

?

slide14

SAM Utilization

Met

MetK

SAM

SAH

MTA

Pfs

Pfs

SRH

MTR

LuxS?

4,5-Dihydroxy-

2,3-pentanedione

+ HC

?

slide15

SAM Utilization

Met

MetK

SAM

SAH

MTA

Pfs

Pfs

SRH

MTR

LuxS

AI-2 + HC

?

slide16

The LuxS Reaction

4,5-Dihydroxy-

2,3-pentanedione

SRH

+

Homocysteine

slide18

LuxP

LuxR

LuxCDABE

Quorum Sensing in V. harveyi

AI-2

LuxQ

H

D

H

D

HTH

LuxU

LuxO

H

D

LuxN

AI-1

slide23

Boron is ubiquitous in the biosphere

Sea water 0.4 mM (primarily boric acid)

L.A. tap water 40 μM

Soils 2-100 ppm

Marine plants 120 ppm

Land plants 50 ppm

Marine animals 20-50 ppm

Land animals 0.5 ppm

Boron is required by a number of organisms,

but for poorly characterized reasons

Vascular plants cell wall? pollination?

Diatoms

Cyanobacteria heterocyst formation

Zebrafish embryogenesis

slide24

11BNMR

holo-LuxP

filtered

apo-LuxP

slide28

Targets of AI-2

Vibrio harveyi Bioluminescence, Biofilms

Vibrio cholerae Virulence, Biofilms

Salmonella typhimurium ABC transporter

Salmonella typhi Biofilms

Escherichia coli O157:H7Type III Secretion (virulence)

Porphyromonas gingivalis Hemin acquisition genes, virulence factors

Shigella flexneri VirB (virulence factor)

Streptococcus pyogenes Secreted protease

Actinobacillus actinomycetemcomitans Periodontal virulence, Mixed-species biofilms

Photorhabdus luminescens Antibiotic production

Neisseria meningitidis Virulence/bacteremia

Borrelia burgdorferi Virulence

Clostridium perfringens Virulence

Campylobacter jejuni Motility

Microarrays

Escherichia coli K12 ~300 genes

Escherichia coli O157:H7 ~400 genes

Vibrio cholerae El Tor ~70 genes (virulence regulon)

slide29

Biosynthesis of AI-1 and AI-2

Met

MetK

LuxI

SAM

LuxLM

AI-1

SAH

MTA

Pfs

Pfs

SRH

MTR

LuxS

AI-2 + HC

?

slide30

LuxP

LuxR

LuxCDABE

Quorum Sensing in V. harveyi

AI-2

LuxQ

H

D

H

D

HTH

LuxU

LuxO

H

D

LuxN

AI-1