Bacillus subtilis & Myxococcus xanthus

Intra- & Inter-species communication

& Predation

Bacillus subtilis

Myxococcus xanthus

Bacillus subtilis & Myxococcus xanthus

Intra- & Inter-species communication
& Predation

comparison of B. subtilis & M. xanthus

B. subtilis biofilm formation
B. subtilis endospore formation
M. xanthus fruiting body formation

Inter-species communication
Bacterial predation

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites

GC content: 43.5%
4.2 Mbp genome

GC content: 68.9%
9.1 Mbp genome

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites

GC content: 43.5%
4.2 Mbp genome

GC content: 68.9%
9.1 Mbp genome

motility: flagella

focal adhesion/T4P

Myxococcus motility: pili & focal adhesion complexes

Focal adhesion complex mediated motility

MreB actin-like protein forms helical

filaments
AglZ motility protein forms focal adhesion
Pili are retractable appendages
PilA is the major component of pilus fiber
PilT ATPase = pilus retraction motor

complexes that are attached to MreB

filaments

Hypothetical motor interacts with focal

adhesion complexes & produce trust
tracking on the internal cytoskeleton
filaments

Mauriello et al, 2011

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites
GC content: 43.5%
4.2 Mbp genome

GC content: 68.9%
9.2 Mbp genome

motility: flagella

focal adhesion/T4P

gram-positive

gram-negative

Bacterial cell wall & gram staining

Hans Christian Gram (1884)

differentiate bacteria by chemical

& physical properties
detects peptidoglycan
Gram-positive: purple
Gram-negative: pink-red

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites

GC content: 43.5%
4.2 Mbp genome

GC content: 68.9%
9.2 Mbp genome

motility: flagella

focal adhesion/T4P

gram-positive

gram-negative

Endospore formation

Spore formation

Myxococcus xanthus spores

Resistant to heat, UV, sonication,

chemical stresses

Whole cell converts into a spore

Spore coat material carbohydraterich (1-4,1-3-linked GalNAc)
Multilayer spore coat on surface of
OM
Lipid bodies provide metabolic
fuel (intermediate carbon &
energy storage)

Bacillus subtilis sporulation cycle

Kin Kinases - master regulator

Spo0A-P (about 120 genes)

Intercompartmental signalling
Temporal, spatial separation
Low water content, low ph

Dipicolinic acid, SASP protect

DNA
Germination (sugars, amino
acids, PG)

McKenney et al., 2013

Bacillus subtilis spore layers

Cortex: peptidoglycan
Coat: Basement layer, Inner & outer coat,
crust (70 individual proteins)

McKenney et al., 2013

Bacillus subtilis sporulation control by sigma factors

Genetic asymmetry
Spo0A-P & Sigma H trigger
division
Sigma F & G in forespore
Sigma E & K in mother cell

Intercelluar signaling cascade

SpoIIR (Sigma F control)
? factor enters forespore &
activates Sigma G
SpoIVB activates Sigma K

Higgins & Dworkin., 2012

Bacillus subtilis biofilm formation

Vlamakis et al., 2013

Bacillus subtilis biofilm signaling network

Components of biofilm matrix

EPS
TasA amyloid fibers
BslA (biofilm surface layer
protein, hydrophobic layer)
PGS (glutamic acid)

Regulators of biofilm formation

DegS - DegU-P (regulator of
motility, competence, matrix)
Surfactin (paracrine signaling)
Spo0A activated by Kin kinases
SinR master regulator, repressor
of matrix production
SinI anti-repressor of SinR
Spo0A-P low: SinI expressed
Spo0A-P high: SinI concentration
low, SinR derepression

Vlamakis et al., 2013

Myxococcus fruiting body formation

Nutrient deprivation sensing (stringent response)

A signaling (early in development, amino acids)
C signaling (later in development, accumulates on cell
surface)
Complex signaling cascades (Two-component systems)
Motility

Kuner & Kaiser, 1982

Bacillus subtilis & Myxococcus xanthus

soil bacteria
multicellular behavior
Inter-& Intra-species
interactions
secondary metabolites

GC content: 43.5%
4.2 Mbp genome

GC content: 68.9%
9.2 Mbp genome

motility: flagella

focal adhesion/T4P

gram-positive

gram-negative

Endospore formation

Spore formation
Predator

The Importance of Predation

Global Diversity
Population control
Shaping community structure
(diversity, dynamics)

Biogeochemical Diversity
Degradation of polymeric compounds
Control of nutrient availability
& recycling

Host Specific Interactions

Predator-Prey interactions can be specialized
Competition
Intraspecies Interactions

Predation assays
Standard
Prey

Myxo DZ2

Inside out

Spread plating

Ancestral B. subtilis strain NCIB3610 resists

predation
Predator

E. coli

B. subtilis
168

B. subtilis
NCIB3610

buffer

48 hours

Mller et al., 2014 AEM (80): 5603-10

Bacillaene Protects Bacillus subtilis from Predation

Bacillaene (pks)

Inside-out Predation assay

pksL 4% survival

Mller et al., 2014 AEM (80): 5603-10

Spores Provide the Ultimate Protection Against

Predation
vegetative cells

spores

laboratory
strains

NCIB3610

NCIB3610 pksL
(bacillaene-)

Mller et al., 2014 AEM (80): 5603-10

Megastructures Appear During Long-Term Predation

Assays

lab strain of B.
subtilis

B. subtilis
NCIB3610

Megastructure

Mller et al., 2014 AEM

Megastructures Contain Spores & Matrix

Mller et al., 2014 AEM

Megastructures Form On or Near Fruiting Bodies

Mller et al., 2014 AEM

Bacillus

Myxococcus

Fruiting Body-Megastructure Intersection

Mller et al., 2014 AEM

Spore Completion is Required for Structure Integrity

NCIB3610

spoIVA

Mller et al., 2014 AEM

Bacillus subtilis biofilm signalling network

Vlamakis et al., 2013

Megastructures are Genetically Distinguished from

B. subtilis Colony Biofilm Formation
MSgg
A

predator
B -
C+

MSgg
A

NCIB3610

sinI

epsE

sinR

tasA

degU

epsE
tasA

spoIVA

bslA

spo0A

predator
B -
C +

Mller et al., 2014 AEM

Myxo-Bacillus Interaction Model

lytic enzymes, T4P

secondary metabolites

predation-induced
stress response

Bacillaene

Myxococcus
xanthus

Bacillus subtilis
NCIB 3610

B. subtilis
megastructure
(sporulation)

Birds and the Bees: Mites eating & transporting fruiting bodies?

Susanne Mller