Global Control of Aspergillus fumigatus Toxins

Nancy Keller

University of Wisconsin

Aspergillus secondary metabolites

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Aspergillus laeA: transcriptional regulator of secondary metabolite genes

Bok and Keller, Euk Cell, 3: 527-535

ST WT laeA

A. nidulans

GT WT laeA comp

A. fumigatus

Time (Days)0 2 4 6 8

0

20

40

60

80

100

laeAWild type

0 2 4 6 8

20

40

60

80

100

0 2 4 6 8

20

40

60

80

100

Sur

viva

l %

0 2 4 6 8

20

40

60

80

100

Complemented

Mouse survival

LaeA is a virulence factor in Aspergillus fumigatus

Bok et al, Euk Cell, 4:1574-15825 15 60

0

10

20

30

40

50

60 WTlaeA

Incubation (minutes)

PM

N d

eath

(%

)

Macrophage phagocytosis

WT laeA

Neutrophil death

GObiologicalprocesses GO

#genes down up

high-down mod-down high-up mod-up

alldlaeA genome

signaltransduction I 6 (0) 2 4 0 2 1 3 6 133lipidmetabolism II 8 (0) 3 5 2 1 2 3 8 354response tostress III 10 (2) 6 (2) 4 4 2 0 4 10 213carbohydratemetabolism IV 15 (4) 7 (3)

8(1) 3 4 3 5 15 294

cell wallbiogenesis V 19 (0) 10 9 7 3 5 4 19 176regulation oftranscription VI 20 (1) 9 (1) 11 5 4 3 8 20 388

amino acid VII 21 (1) 8 (1) 14 3 5 10 4 22 197cellularmetabolism VIII 22 (3) 11 (3) 12 7 4 4 8 23 2000

transport IX 35 (1) 11 (1) 23 6 5 12 11 34 603secondarymetabolism X

90(88)

87(87)

2(1) 72 15 2 0 89 224

unknown XI144(8) 72 (4)

72(4) 47 25 21 51 144 4000

total total409

(108)232

(102)173(6) 159 73 69 104 405 9858

Gardiner and Howlett 2005, Microbiology

Nu

mbe

r of

con

idia

(1

06/m

l/cm

2)

0

10

20

30

40

50

60

WT gliZ gliZ + gliZ gliZ + gliZ

A. fumigatus gliZ grows at same rate, produces same number of

spores

Gt WT gliZ gliZ + gliZ

gliZ + gliZ

laeA

gliZ does not produce gliotoxin

gliZ regulates other genes in the gliotoxin gene cluster

gliZ

rRNA

36hr 72hr

WT laeA gliZ gliZcom WT laeA gliZ gliZcom

gliI

A B

0.5 kb

1.0 kb

1 2 3 4

Murine pulmonary model

Time (Days post infection)

0 2 4 6 8

0

20

40

60

80

100

0 2 4 6 8

0

20

40

60

80

100

0 2 4 6 8

0

20

40

60

80

100

Sur

viva

l (%

)

0 2 4 6 8

0

20

40

60

80

100

WT

laeA

gliZ

gliZcom8.3

Gliotoxinstandard

Non-infected lung

gliZ

laeA

WT

1000

5.00 5.20 5.40 5.60 5.80 6.00 6.20 6.40 6.60 6.80 7.00 7.20Time (min)

0

0

17

0

520

0

19

0

312

0

925.87

5.80

5.87

5.87

Gliotoxin chromatograms lung tissue

gliZ + gliZ

A

B p=0.04

p=0.04

0

5

10

15

20

25

30

35

40

45

WT laeA gliZ gliZcom8.3

PI

+ (

% P

MN

s)

0

10

20

30

40

50

60

An

nex

in +

(%

PM

Ns)

WT laeA gliZ gliZcom8.3

Acknowledgements• Keller Lab

– Dr. JinWoo Bok– Elyse Bolterstein– Dr. Marion Brodhagen– DaWoon Chung– Tom Hammond– Lane Milde– Dr. Robyn Perrin– James Scott– E. Keats Shwab

• Collaborators– Drs. Bill Nierman, Natalie Fedorova

• The Institute for Genomic Research (microarrays)The Institute for Genomic Research (microarrays)– Dr. David Andes

• Department of Medicine, Section of Infectious Disease, UW Madison

– Drs. Kieren Marr and Arunmozhi Balajee• Fred Hutchinson Cancer Research Center

– Drs. Jens Frisvad & Kristian Fog Nielsen• Department of Biotechnology, Technical University of

Denmark