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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
A. fumigatus
A. nidulans
ST WT
laeA
GT WT laeA comp
Bok and Keller, Euk Cell, 3: 527-535
LaeA is a virulence factor in Aspergillus fumigatus
Macrophage phagocytosis
Mouse survival
80
laeA
60
WT
40
Neutrophil death
laeA
20
Wild type
Complemented
0
0
2
4
6
Time (Days)
8
PMN death (%)
Survival %
10
100
0
60
WT
laeA
50
40
30
20
10
0
Bok et al, Euk Cell, 4:1574-1582
5
15
60
Incubation (minutes)
GO
biological
proce sse s
signal
transduction
lipid
me tabolism
response to
stres s
carbohydrate
me tabolism
cell wall
bioge nesis
re gulation of
transcription
amino acid
cellular
me tabolism
transport
secondary
me tabolism
unknown
total
GO
#
genes
down
up
highdown
mod-down
high-up
mod-up
all
dlaeA
genome
I
6 (0)
2
4
0
2
1
3
6
133
II
8 (0)
3
5
2
1
2
3
8
354
III
10 (2)
6 (2)
4
2
0
4
10
213
IV
15 (4)
7 (3)
4
8
(1)
3
4
3
5
15
294
V
19 (0)
10
9
7
3
5
4
19
176
VI
VII
20 (1)
21 (1)
9 (1)
8 (1)
11
14
5
3
4
5
3
10
8
4
20
22
388
197
VIII
IX
22 (3)
35 (1)
90
(88)
144
(8)
409
(108)
11 (3)
11 (1)
87
(87)
12
23
2
(1)
72
(4)
173
(6)
7
6
4
5
4
12
8
11
23
34
2000
603
72
15
2
0
89
224
47
25
21
51
144
4000
159
73
69
104
405
9858
X
XI
total
72 (4)
232
(102)
Gardiner and Howlett 2005, Microbiology
Number of conidia (106/ml/cm2)
A. fumigatus gliZ grows at same
rate, produces same number of
spores
60
50
40
30
20
10
0
WT
gliZ
gliZ + gliZ gliZ + gliZ
gliZ does not produce gliotoxin
Gt
laeA
WT
gliZ
gliZ
gliZ
+ gliZ
+ gliZ
gliZ regulates other genes in the gliotoxin gene
cluster
B
A
1 2 3 4
36hr
WT laeA gliZ
72hr
gliZcom
WT laeA gliZ
gliZcom
gliZ
gliI
1.0 kb
0.5 kb
rRNA
Murine pulmonary model
100
Survival (%)
80
60
40
WT
laeA
20
gliZ
gliZcom8.3
0
0
2
4
6
Time (Days post infection)
8
Gliotoxin chromatograms lung tissue
5.87
92
laeA
0
5.80
312
WT
0
19
gliZ
0
5.87
520
gliZ + gliZ
0
Non-infected
lung
Gliotoxin
standard
17
0
5.87
1000
0
5.00
5.20
5.40
5.60
5.80
6.00 6.20 6.40
Time (min)
6.60
6.80
7.00
7.20
PI + (% PMNs)
A
45
40
35
30
25
20
15
10
5
0
WT
laeA
gliZ gliZcom8.3
p=0.04
B
p=0.04
Annexin + (% PMNs)
60
50
40
30
20
10
0
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)
– 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