global control of aspergillus fumigatus toxins nancy keller university of wisconsin
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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
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